Prodrug modulators of the integrated stress pathway

ABSTRACT

Provided herein are compounds, compositions, and methods useful for modulating the integrated stress response (ISR) and for treating related diseases, disorders and conditions.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No. 17/227,071, filed on Apr. 9, 2021, now issued as U.S. Pat. No. 11,149,043 on Oct. 19, 2021, which is a continuation of International Patent Application No. PCT/US2019/055850, filed on Oct. 11, 2019, which claims the benefit of, and priority to, U.S. Provisional Application No. 62/744,293, filed on Oct. 11, 2018, the contents of each of which are hereby incorporated by reference in their entirety.

BACKGROUND

In metazoa, diverse stress signals converge at a single phosphorylation event at serine 51 of a common effector, the translation initiation factor eIF2α. This step is carried out by four eIF2α kinases in mammalian cells: PERK, which responds to an accumulation of unfolded proteins in the endoplasmic reticulum (ER), GCN2 to amino acid starvation and UV light, PKR to viral infection and metabolic stress, and HRI to heme deficiency. This collection of signaling pathways has been termed the “integrated stress response” (ISR), as they converge on the same molecular event. eIF2α phosphorylation results in an attenuation of translation with consequences that allow cells to cope with the varied stresses (Wek, R. C. et al, Biochem Soc Trans (2006) 34(Pt 1):7-11).

eIF2 (which is comprised of three subunits, α, β and γ) binds GTP and the initiator Met-tRNA to form the ternary complex (eIF2-GTP-Met-tRNA), which, in turn, associates with the 40S ribosomal subunit scanning the 5′UTR of mRNAs to select the initiating AUG codon. Upon phosphorylation of its α-subunit, eIF2 becomes a competitive inhibitor of its GTP-exchange factor (GEF), eIF2B (Hinnebusch, A. G. and Lorsch, J. R. Cold Spring Harbor Perspect Biol (2012) 4(10)). The tight and nonproductive binding of phosphorylated eIF2 to eIF2B prevents loading of the eIF2 complex with GTP, thus blocking ternary complex formation and reducing translation initiation (Krishnamoorthy, T. et al, Mol Cell Biol (2001) 21(15):5018-5030). Because eIF2B is less abundant than eIF2, phosphorylation of only a small fraction of the total eIF2 has a dramatic impact on eIF2B activity in cells.

eIF2B is a complex molecular machine, composed of five different subunits, eIF2B1 through eIF2B5. eIF2B5 catalyzes the GDP/GTP exchange reaction and, together with a partially homologous subunit eIF2B3, constitutes the “catalytic core” (Williams, D. D. et al, J Biol Chem (2001) 276:24697-24703). The three remaining subunits (eIF2B1, eIF2B2, and eIF2B4) are also highly homologous to one another and form a “regulatory sub-complex” that provides binding sites for eIF2B's substrate eIF2 (Dev, K. et al, Mol Cell Biol (2010) 30:5218-5233). The exchange of GDP with GTP in eIF2 is catalyzed by its dedicated guanine nucleotide exchange factor (GEF) eIF2B. eIF2B exists as a decamer (B1₂B2₂ B3₂ B4₂ B5₂) or dimer of two pentamers in cells (Gordiyenko, Y. et al, Nat Commun (2014) 5:3902; Wortham, N.C. et al, FASEB J (2014) 28:2225-2237). Molecules such as ISRIB interact with and stabilize the eIF2B dimer conformation, thereby enhancing intrinsic GEF activity and making cells less sensitive to the cellular effects of phosphorylation of eIF2α (Sidrauski, C. et al, eLife (2015) e07314; Sekine, Y. et al, Science (2015) 348:1027-1030). As such, small molecule therapeutics that can modulate eIF2B activity may have the potential to attenuate the PERK branch of the UPR and the overall ISR, and therefore may be used in the prevention and/or treatment of various diseases, such as a neurodegenerative disease, a leukodystrophy, a cancer, an inflammatory disease, an autoimmune disease, a viral infection, a skin disease, a fibrotic disease, a hemoglobin disease, a kidney disease, a hearing loss condition, an ocular disease, a musculoskeletal disease, or a metabolic disease.

SUMMARY OF THE INVENTION

In one aspect, described herein is a compound represented by Formula (I):

or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, wherein:

R¹ is selected from the group consisting of —C(O)—C₁₋₄alkyl, —C(O)—O—C₁₋₄alkyl, —C(O)—N(R^(a))—C₁₋₄alkyl, —C(O)—C₁₋₄alkylene-C₁₋₄alkoxy, —C(O)—C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, -methylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—O—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—N(R^(a))—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-P(O)(OH)₂, —C(O)—C₁₋₅ alkylene-phenylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—C₁₋₅ alkylene-phenylene-(O—P(O)(OH)₂)(O—C₁₋₅alkylene-P(O)(OH)₂), -methylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, -methylene-O—C(O)C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—O—C₁₋₅alkylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, —C(O)—N(R^(a))-heteroarylene-C₁₋₂alkylene-O—P(O)(OH)₂, —P(O)(OH)₂, —SO₃H, —SO₂NR^(a)R^(b), —C(O)-heteroaryl, —C(O)—C₁₋₅alkylene-O—C₁₋₅alkylene-phenyl and methylene-C₁₋₅alkoxide;

wherein —C(O)—C₁₋₄alkyl is substituted by one or two substituents each independently selected from the group consisting of NR^(a)R^(b) and —CO₂H; and wherein —C(O)—O—C₁₋₄alkyl, —C(O)—N(R^(a))—C₁₋₄alkyl, —C(O)—C₁₋₄alkylene-C₁₋₄alkoxy, —C(O)—C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, methylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—O—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—N(R^(a))—C₁₋₅ alkylene-O—P(O)(OH)₂, —C(O)—C₁₋₅ alkylene-P(O)(OH)₂, —C(O)—C₁₋₅ alkylene-phenylene-O—P(O)(OH)₂, —C(O)—C₁₋₅ alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—C₁₋₅ alkylene-phenylene-(O—P(O)(OH)₂)(O—C₁₋₅ alkylene-P(O)(OH)₂), -methylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, -methylene-O—C(O)C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—O—C₁₋₅alkylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, —C(O)-heteroaryl, —C(O)—C₁₋₅alkylene-O—C₁₋₅alkylene-phenyl and —C(O)—N(R^(a))-heteroarylene-C₁₋₂alkylene-O—P(O)(OH)₂ may optionally be substituted by one, two, three or four substituents each independently selected from the group consisting of halogen, —CO₂H, —NR^(a)R^(b) and C₁₋₂alkyl (optionally substituted by one, two or three fluorines) and aryl; and

R^(a) and R^(b) are independently selected, for each occurrence, from the group consisting of hydrogen and C₁₋₃alkyl.

In one aspect, described herein is a pharmaceutically acceptable composition comprising any one of the compounds disclosed herein and a pharmaceutically acceptable carrier.

In one aspect, described herein is a method of treating a neurodegenerative disease, a leukodystrophy, a cancer, an inflammatory disease, an autoimmune disease, a viral infection, a skin disease, a fibrotic disease, a hemoglobin disease, a kidney disease, a hearing loss condition, an ocular disease, a musculoskeletal disease, a metabolic disease, or a mitochondrial disease in a patient in need thereof, comprising administering to the patient an effective amount of any one of the compounds disclosed herein.

In one aspect, described herein is a method of treating a disease related to a modulation of eIF2B activity or levels, eIF2α activity or levels, or the activity or levels of a component of the eIF2 pathway or the ISR pathway in a patient in need thereof, comprising administering to the patient an effective amount of any one of the compounds disclosed herein.

In one aspect, described herein is a method of treating a cancer in a patient in need thereof, comprising administering to the patient any one of the compounds disclosed herein in combination with an immunotherapeutic agent.

DETAILED DESCRIPTION OF THE INVENTION

The present invention features compounds, compositions, and methods comprising a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof for use, e.g., in the modulation (e.g., activation) of eIF2B and the attenuation of the ISR signaling pathway. In some embodiments, a compound of Formula (I) is a prodrug of a biologically active compound that modulates eIF2B, eIF2α, or a component of the eIF2 or ISR pathways. In some embodiments, a compound of Formula (I) is a prodrug of the compound of Formula (II):

Definitions Chemical Definitions

Definitions of specific functional groups and chemical terms are described in more detail below. The chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75^(th) Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, 5^(th) Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987.

The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various isomeric forms, e.g., enantiomers and/or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind. 1972). The invention additionally encompasses compounds described herein as individual isomers substantially free of other isomers, and alternatively, as mixtures of various isomers.

As used herein a pure enantiomeric compound is substantially free from other enantiomers or stereoisomers of the compound (i.e., in enantiomeric excess). In other words, an “S” form of the compound is substantially free from the “R” form of the compound and is, thus, in enantiomeric excess of the “R” form. The term “enantiomerically pure” or “pure enantiomer” denotes that the compound comprises more than 75% by weight, more than 80% by weight, more than 85% by weight, more than 90% by weight, more than 91% by weight, more than 92% by weight, more than 93% by weight, more than 94% by weight, more than 95% by weight, more than 96% by weight, more than 97% by weight, more than 98% by weight, more than 99% by weight, more than 99.5% by weight, or more than 99.9% by weight, of the enantiomer. In certain embodiments, the weights are based upon total weight of all enantiomers or stereoisomers of the compound.

In the compositions provided herein, an enantiomerically pure compound can be present with other active or inactive ingredients. For example, a pharmaceutical composition comprising enantiomerically pure R-compound can comprise, for example, about 90% excipient and about 10% enantiomerically pure R-compound. In certain embodiments, the enantiomerically pure R compound in such compositions can, for example, comprise, at least about 95% by weight R compound and at most about 5% by weight S-compound, by total weight of the compound. For example, a pharmaceutical composition comprising enantiomerically pure S-compound can comprise, for example, about 90% excipient and about 10% enantiomerically pure S-compound. In certain embodiments, the enantiomerically pure S-compound in such compositions can, for example, comprise, at least about 95% by weight S-compound and at most about 5% by weight R-compound, by total weight of the compound. In certain embodiments, the active ingredient can be formulated with little or no excipient or carrier.

Compounds described herein may also comprise one or more isotopic substitutions. For example, H may be in any isotopic form, including ¹H, ²H (D or deuterium), and ³H (T or tritium); C may be in any isotopic form, including ¹²C, ¹³C, and ¹⁴C; O may be in any isotopic form, including ¹⁶O and ¹⁸O; and the like.

The articles “a” and “an” may be used herein to refer to one or to more than one (i.e. at least one) of the grammatical objects of the article. By way of example “an analogue” means one analogue or more than one analogue.

When a range of values is listed, it is intended to encompass each value and sub-range within the range. For example “C₁-C₆ alkyl” is intended to encompass, C₁, C₂, C₃, C₄, C₅, C₆, C₁-C₆, C₁-C₅, C₁-C₄, C₁-C₃, C₁-C₂, C₂-C₆, C₂-C₅, C₂-C₄, C₂-C₃, C₃-C₆, C₃-C₅, C₃-C₄, C₄-C₆, C₄-C₅, and C₅-C₆ alkyl.

The following terms are intended to have the meanings presented therewith below and are useful in understanding the description and intended scope of the present invention.

“Alkyl” refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 20 carbon atoms (“C₁-C₂₀ alkyl”). In some embodiments, an alkyl group has 1 to 12 carbon atoms (“C₁-C₁₂ alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C₁-C₈ alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C₁-C₆ alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C₁-C₅ alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C₁-C₄alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C₁-C₃ alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C₁-C₂ alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C₁ alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C₂-C₆alkyl”). Examples of C₁-C₆alkyl groups include methyl (C₁), ethyl (C₂), n-propyl (C₃), isopropyl (C₃), n-butyl (C₄), tert-butyl (C₄), sec-butyl (C₄), iso-butyl (C₄), n-pentyl (C₅), 3-pentanyl (C₅), amyl (C₅), neopentyl (C₅), 3-methyl-2-butanyl (C₅), tertiary amyl (C₅), and n-hexyl (C₆). Additional examples of alkyl groups include n-heptyl (C₇), n-octyl (C₈) and the like. Each instance of an alkyl group may be independently optionally substituted, i.e., unsubstituted (an “unsubstituted alkyl”) or substituted (a “substituted alkyl”) with one or more substituents; e.g., for instance from 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkyl group is unsubstituted C₁₋₁₀ alkyl (e.g., —CH₃). In certain embodiments, the alkyl group is substituted C₁₋₆ alkyl. Common alkyl abbreviations include Me (—CH₃), Et (—CH₂CH₃), iPr (—CH(CH₃)₂), nPr (—CH₂CH₂CH₃), n-Bu (—CH₂CH₂CH₂CH₃), or i-Bu (—CH₂CH(CH₃)₂).

The term “alkylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkyl, as exemplified, but not limited by, —CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being preferred in the present invention. The term “alkenylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkene. An alkylene group may be described as, e.g., a C₁-C₆-membered alkylene, wherein the term “membered” refers to the non-hydrogen atoms within the moiety.

“Alkenyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 20 carbon atoms, one or more carbon-carbon double bonds, and no triple bonds (“C₂-C₂₀ alkenyl”). In some embodiments, an alkenyl group has 2 to 10 carbon atoms (“C₂-C₁₀ alkenyl”). In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C₂-C₅ alkenyl”). In some embodiments, an alkenyl group has 2 to 6 carbon atoms (“C₂-C₆ alkenyl”). In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C₂-C₅ alkenyl”). In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C₂-C₄ alkenyl”). In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C₂-C₃ alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C₂ alkenyl”). The one or more carbon carbon double bonds can be internal (such as in 2butenyl) or terminal (such as in 1butenyl). Examples of C₂-C₄ alkenyl groups include ethenyl (C₂), 1propenyl (C₃), 2propenyl (C₃), 1 butenyl (C₄), 2butenyl (C₄), butadienyl (C₄), and the like. Examples of C₂-C₆ alkenyl groups include the aforementioned C₂₋₄ alkenyl groups as well as pentenyl (C₅), pentadienyl (C₅), hexenyl (C₆), and the like. Additional examples of alkenyl include heptenyl (C₇), octenyl (C₈), octatrienyl (C₈), and the like. Each instance of an alkenyl group may be independently optionally substituted, i.e., unsubstituted (an “unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) with one or more substituents e.g., for instance from 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. In certain embodiments, the alkenyl group is unsubstituted C₂₋₁₀ alkenyl. In certain embodiments, the alkenyl group is substituted C₂₋₆ alkenyl.

“Aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 it electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C₆-C₁₄ aryl”). In some embodiments, an aryl group has six ring carbon atoms (“C₆ aryl”; e.g., phenyl). In some embodiments, an aryl group has ten ring carbon atoms (“C₁₀ aryl”; e.g., naphthyl such as 1 naphthyl and 2naphthyl). In some embodiments, an aryl group has fourteen ring carbon atoms (“C₁₄ aryl”; e.g., anthracyl). An aryl group may be described as, e.g., a C₆-C₁₀-membered aryl, wherein the term “membered” refers to the non-hydrogen ring atoms within the moiety. Aryl groups include, but are not limited to, phenyl, naphthyl, indenyl, and tetrahydronaphthyl. Each instance of an aryl group may be independently optionally substituted, i.e., unsubstituted (an “unsubstituted aryl”) or substituted (a “substituted aryl”) with one or more substituents. In certain embodiments, the aryl group is unsubstituted C₆-C₁₄ aryl. In certain embodiments, the aryl group is substituted C₆-C₁₄ aryl.

In certain embodiments, an aryl group is substituted with one or more of groups selected from halo, C₁-C₈ alkyl, halo-C₁-C₈ alkyl, haloxy-C₁-C₈ alkyl, cyano, hydroxy, alkoxy C₁-C₈ alkyl, and amino.

Examples of representative substituted aryls include the following

wherein one of R⁵⁶ and R⁵⁷ may be hydrogen and at least one of R⁵⁶ and R⁵⁷ is each independently selected from C₁-C₈ alkyl, halo-C₁-C₈ alkyl, 4-10 membered heterocyclyl, alkanoyl, alkoxy-C₁-C₈ alkyl, heteroaryloxy, alkylamino, acylamino, heteroarylamino, NR⁵⁸COR⁵⁹, NR⁵⁸SOR⁵⁹NR⁵⁸SO₂R⁵⁹, C(O)Oalkyl, C(O)Oaryl, CONR⁵⁸R⁵⁹, CONR⁵⁸OR⁵⁹, NR⁵⁸R⁵⁹, SO₂NR⁵⁸R⁵⁹, S-alkyl, S(O)-alkyl, S(O)₂-alkyl, S-aryl, S(O)-aryl, S(O₂)-aryl; or R⁵⁶ and R⁵⁷ may be joined to form a cyclic ring (saturated or unsaturated) from 5 to 8 atoms, optionally containing one or more heteroatoms selected from the group N, O, or S.

Other representative aryl groups having a fused heterocyclyl group include the following:

wherein each W′ is selected from C(R⁶⁶)₂, NR⁶⁶, O, and S; and each Y′ is selected from carbonyl, NR⁶⁶, O and S; and R⁶⁶ is independently hydrogen, C₁-C₈ alkyl, C₃-C₁₀cycloalkyl, 4-10 membered heterocyclyl, C₆-C₁₀aryl, and 5-10 membered heteroaryl.

An “arylene” and a “heteroarylene,” alone or as part of another substituent, mean a divalent radical derived from an aryl and heteroaryl, respectively. Non-limiting examples of heteroaryl groups include pyridinyl, pyrimidinyl, thiophenyl, thienyl, furanyl, indolyl, benzoxadiazolyl, benzodioxolyl, benzodioxanyl, thianaphthanyl, pyrrolopyridinyl, indazolyl, quinolinyl, quinoxalinyl, pyridopyrazinyl, quinazolinonyl, benzoisoxazolyl, imidazopyridinyl, benzofuranyl, benzothienyl, benzothiophenyl, phenyl, naphthyl, biphenyl, pyrrolyl, pyrazolyl, imidazolyl, pyrazinyl, oxazolyl, isoxazolyl, thiazolyl, furylthienyl, pyridyl, pyrimidyl, benzothiazolyl, purinyl, benzimidazolyl, isoquinolyl, thiadiazolyl, oxadiazolyl, pyrrolyl, diazolyl, triazolyl, tetrazolyl, benzothiadiazolyl, isothiazolyl, pyrazolopyrimidinyl, pyrrolopyrimidinyl, benzotriazolyl, benzoxazolyl, or quinolyl. The examples above may be substituted or unsubstituted and divalent radicals of each heteroaryl example above are non-limiting examples of heteroarylene.

“Halo” or “halogen,” independently or as part of another substituent, mean, unless otherwise stated, a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom. The term “halide” by itself or as part of another substituent, refers to a fluoride, chloride, bromide, or iodide atom. In certain embodiments, the halo group is either fluorine or chlorine.

Additionally, terms such as “haloalkyl” are meant to include monohaloalkyl and polyhaloalkyl. For example, the term “halo-C₁-C₆ alkyl” includes, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “heteroalkyl,” by itself or in combination with another term, means, unless otherwise stated, a non-cyclic stable straight or branched chain, or combinations thereof, including at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si, and S, and wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) O, N, P, S, and Si may be placed at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Exemplary heteroalkyl groups include, but are not limited to: —CH₂—CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃, —CH₂—CH₂, —S(O)₂, —S(O)—CH₃, —S(O)₂—CH₂, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃, —CH₂—CH═N—OCH₃, —CH═CH—N(CH₃)—CH₃, —O—CH₃, and —O—CH₂—CH₃. Up to two or three heteroatoms may be consecutive, such as, for example, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃. Where “heteroalkyl” is recited, followed by recitations of specific heteroalkyl groups, such as CH₂O, NR^(B)R^(C), or the like, it will be understood that the terms heteroalkyl and CH₂O or NR^(B)R^(C) are not redundant or mutually exclusive. Rather, the specific heteroalkyl groups are recited to add clarity. Thus, the term “heteroalkyl” should not be interpreted herein as excluding specific heteroalkyl groups, such as CH₂O, NR^(B)R^(C), or the like.

Similarly, the term “heteroalkylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from heteroalkyl, as exemplified, but not limited by, CH₂O— and CH₂CH₂O—. A heteroalkylene group may be described as, e.g., a 2-7-membered heteroalkylene, wherein the term “membered” refers to the non-hydrogen atoms within the moiety. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like). Still further, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, the formula —C(O)₂R′— may represent both —C(O)₂R′— and R′C(O)₂—.

“Heteroaryl” refers to a radical of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur (“5-10 membered heteroaryl”). In heteroaryl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. Heteroaryl bicyclic ring systems can include one or more heteroatoms in one or both rings. “Heteroaryl” also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused (aryl/heteroaryl) ring system. Bicyclic heteroaryl groups wherein one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and the like) the point of attachment can be on either ring, i.e., either the ring bearing a heteroatom (e.g., 2indolyl) or the ring that does not contain a heteroatom (e.g., 5indolyl). A heteroaryl group may be described as, e.g., a 6-10-membered heteroaryl, wherein the term “membered” refers to the non-hydrogen ring atoms within the moiety.

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-8 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In some embodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Each instance of a heteroaryl group may be independently optionally substituted, i.e., unsubstituted (an “unsubstituted heteroaryl”) or substituted (a “substituted heteroaryl”) with one or more substituents. In certain embodiments, the heteroaryl group is unsubstituted 5-14 membered heteroaryl. In certain embodiments, the heteroaryl group is substituted 5-14 membered heteroaryl.

Exemplary 5membered heteroaryl groups containing one heteroatom include, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary 5membered heteroaryl groups containing two heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5membered heteroaryl groups containing three heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5membered heteroaryl groups containing four heteroatoms include, without limitation, tetrazolyl. Exemplary 6membered heteroaryl groups containing one heteroatom include, without limitation, pyridinyl. Exemplary 6membered heteroaryl groups containing two heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6 membered heteroaryl groups containing three or four heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively. Exemplary 7membered heteroaryl groups containing one heteroatom include, without limitation, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, without limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

Examples of representative heteroaryls include the following formulae:

wherein each Y is selected from carbonyl, N, NR⁶⁵, O, and S; and R⁶⁵ is independently hydrogen, C₁-C₈ alkyl, C₃-C₁₀cycloalkyl, 4-10 membered heterocyclyl, C₆-C₁₀aryl, and 5-10 membered heteroaryl.

“Cycloalkyl” refers to a radical of a nonaromatic cyclic hydrocarbon group having from 3 to 10 ring carbon atoms (“C₃-C₁₀cycloalkyl”) and zero heteroatoms in the nonaromatic ring system. In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms (“C₃-C₈cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C₃-C₆ cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C₃-C₆ cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C₅-C₁₀ cycloalkyl”). A cycloalkyl group may be described as, e.g., a C₄-C₇-membered cycloalkyl, wherein the term “membered” refers to the non-hydrogen ring atoms within the moiety. Exemplary C₃-C₆ cycloalkyl groups include, without limitation, cyclopropyl (C₃), cyclopropenyl (C₃), cyclobutyl (C₄), cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl (C₅), cyclohexyl (C₆), cyclohexenyl (C₆), cyclohexadienyl (C₆), and the like. Exemplary C₃-C₈ cycloalkyl groups include, without limitation, the aforementioned C₃-C₆ cycloalkyl groups as well as cycloheptyl (C₇), cycloheptenyl (C₇), cycloheptadienyl (C₇), cycloheptatrienyl (C₇), cyclooctyl (C₈), cyclooctenyl (C₈), cubanyl (C₈), bicyclo[1.1.1]pentanyl (C₈), bicyclo[2.2.2]octanyl (C₈), bicyclo[2.1.1]hexanyl (C₆), bicyclo[3.1.1]heptanyl (C₇), and the like. Exemplary C₃-C₁₀ cycloalkyl groups include, without limitation, the aforementioned C₃-C₈ cycloalkyl groups as well as cyclononyl (C₉), cyclononenyl (C₉), cyclodecyl (C₁₀), cyclodecenyl (C₁₀), octahydro-1H-indenyl (C₉), decahydronaphthalenyl (C₁₀), spiro[4.5]decanyl (C₁₀), and the like. As the foregoing examples illustrate, in certain embodiments, the cycloalkyl group is either monocyclic (“monocyclic cycloalkyl”) or contain a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic cycloalkyl”) and can be saturated or can be partially unsaturated. “Cycloalkyl” also includes ring systems wherein the cycloalkyl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is on the cycloalkyl ring, and in such instances, the number of carbons continue to designate the number of carbons in the cycloalkyl ring system. Each instance of a cycloalkyl group may be independently optionally substituted, i.e., unsubstituted (an “unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”) with one or more substituents. In certain embodiments, the cycloalkyl group is unsubstituted C₃-C₁₀ cycloalkyl. In certain embodiments, the cycloalkyl group is a substituted C₃-C₁₀ cycloalkyl.

In some embodiments, “cycloalkyl” is a monocyclic, saturated cycloalkyl group having from 3 to 10 ring carbon atoms (“C₃-C₁₀cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms (“C₃-C₈ cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C₃-C₆ cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C₅-C₆ cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C₅-C₁₀ cycloalkyl”). Examples of C₅-C₆ cycloalkyl groups include cyclopentyl (C₅) and cyclohexyl (C₅). Examples of C₃-C₆ cycloalkyl groups include the aforementioned C₅-C₆ cycloalkyl groups as well as cyclopropyl (C₃) and cyclobutyl (C₄). Examples of C₃-C₈ cycloalkyl groups include the aforementioned C₃-C₆ cycloalkyl groups as well as cycloheptyl (C₇) and cyclooctyl (C₈). Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted (an “unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”) with one or more substituents. In certain embodiments, the cycloalkyl group is unsubstituted C₃-C₁₀cycloalkyl. In certain embodiments, the cycloalkyl group is substituted C₃-C₁₀cycloalkyl.

“Heterocyclyl” or “heterocyclic” refers to a radical of a 3 to 10membered non aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“3-10 membered heterocyclyl”). In heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. A heterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”) or a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”), and can be saturated or can be partially unsaturated. Heterocyclyl bicyclic ring systems can include one or more heteroatoms in one or both rings. “Heterocyclyl” also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more cycloalkyl groups wherein the point of attachment is either on the cycloalkyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system. A heterocyclyl group may be described as, e.g., a 3-7-membered heterocyclyl, wherein the term “membered” refers to the non-hydrogen ring atoms, i.e., carbon, nitrogen, oxygen, sulfur, boron, phosphorus, and silicon, within the moiety. Each instance of heterocyclyl may be independently optionally substituted, i.e., unsubstituted (an “unsubstituted heterocyclyl”) or substituted (a “substituted heterocyclyl”) with one or more substituents. In certain embodiments, the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. In certain embodiments, the heterocyclyl group is substituted 3-10 membered heterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 membered nonaromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“5-10 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-8 membered non aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-6 membered nonaromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In some embodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclyl has one ring heteroatom selected from nitrogen, oxygen, and sulfur.

Exemplary 3membered heterocyclyl groups containing one heteroatom include, without limitation, azirdinyl, oxiranyl, thiorenyl. Exemplary 4membered heterocyclyl groups containing one heteroatom include, without limitation, azetidinyl, oxetanyl and thietanyl. Exemplary 5 membered heterocyclyl groups containing one heteroatom include, without limitation, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl and pyrrolyl-2,5dione. Exemplary 5membered heterocyclyl groups containing two heteroatoms include, without limitation, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5membered heterocyclyl groups containing three heteroatoms include, without limitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6membered heterocyclyl groups containing one heteroatom include, without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6membered heterocyclyl groups containing two heteroatoms include, without limitation, piperazinyl, morpholinyl, dithianyl, dioxanyl. Exemplary 6membered heterocyclyl groups containing two heteroatoms include, without limitation, triazinanyl. Exemplary 7membered heterocyclyl groups containing one heteroatom include, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary 8membered heterocyclyl groups containing one heteroatom include, without limitation, azocanyl, oxecanyl and thiocanyl. Exemplary 5membered heterocyclyl groups fused to a C₆ aryl ring (also referred to herein as a 5,6-bicyclic heterocyclic ring) include, without limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, and the like. Exemplary 6membered heterocyclyl groups fused to an aryl ring (also referred to herein as a 6,6-bicyclic heterocyclic ring) include, without limitation, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

Particular examples of heterocyclyl groups are shown in the following illustrative examples:

wherein each W is selected from CR⁶⁷, C(R⁶⁷)₂, NR⁶⁷, O, and S; and each Y is selected from NR⁶⁷, O, and S; and R⁶⁷ is independently hydrogen, C₁-C₈ alkyl, C₃-C₁₀cycloalkyl, 4-10 membered heterocyclyl, C₆-C₁₀aryl, and 5-10membered heteroaryl. These heterocyclyl rings may be optionally substituted with one or more groups selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl (e.g., amido), aminocarbonylamino, aminosulfonyl, sulfonylamino, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, halogen, hydroxy, keto, nitro, thiol, —S-alkyl, —S-aryl, —S(O)-alkyl, —S(O)-aryl, —S(O)₂-alkyl, and —S(O)₂-aryl. Substituting groups include carbonyl or thiocarbonyl which provide, for example, lactam and urea derivatives.

“Nitrogen-containing heterocyclyl” group means a 4- to 7-membered nonaromatic cyclic group containing at least one nitrogen atom, for example, but without limitation, morpholine, piperidine (e.g. 2-piperidinyl, 3-piperidinyl and 4-piperidinyl), pyrrolidine (e.g. 2-pyrrolidinyl and 3-pyrrolidinyl), azetidine, pyrrolidone, imidazoline, imidazolidinone, 2-pyrazoline, pyrazolidine, piperazine, and N-alkyl piperazines such as N-methyl piperazine. Particular examples include azetidine, piperidone and piperazone.

“Amino” refers to the radical —NR⁷⁰R⁷¹, wherein R⁷⁰ and R⁷¹ are each independently hydrogen, C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 membered heterocyclyl, C₆-C₁₀ aryl, and 5-10 membered heteroaryl. In some embodiments, amino refers to NH₂.

“Cyano” refers to the radical CN.

“Hydroxy” refers to the radical OH.

Alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups, as defined herein, are optionally substituted (e.g., “substituted” or “unsubstituted” alkyl, “substituted” or “unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl, “substituted” or “unsubstituted” cycloalkyl, “substituted” or “unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or “substituted” or “unsubstituted” heteroaryl group). In general, the term “substituted”, whether preceded by the term “optionally” or not, means that at least one hydrogen present on a group (e.g., a carbon or nitrogen atom) is replaced with a permissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction. Unless otherwise indicated, a “substituted” group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position. The term “substituted” is contemplated to include substitution with all permissible substituents of organic compounds, such as any of the substituents described herein that result in the formation of a stable compound. The present invention contemplates any and all such combinations in order to arrive at a stable compound. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety.

Two or more substituents may optionally be joined to form aryl, heteroaryl, cycloalkyl, or heterocyclyl groups. Such so-called ring-forming substituents are typically, though not necessarily, found attached to a cyclic base structure. In one embodiment, the ring-forming substituents are attached to adjacent members of the base structure. For example, two ring-forming substituents attached to adjacent members of a cyclic base structure create a fused ring structure. In another embodiment, the ring-forming substituents are attached to a single member of the base structure. For example, two ring-forming substituents attached to a single member of a cyclic base structure create a spirocyclic structure. In yet another embodiment, the ring-forming substituents are attached to non-adjacent members of the base structure.

A “counterion” or “anionic counterion” is a negatively charged group associated with a cationic quaternary amino group in order to maintain electronic neutrality. Exemplary counterions include halide ions (e.g., F⁻, Cl⁻, Br⁻, I⁻), NO₃ ⁻, ClO₄ ⁻, OH⁻, H₂PO₄ ⁻, HSO₄ ⁻, sulfonate ions (e.g., methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10camphor sulfonate, naphthalene-2sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonic acid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate, ethanoate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, and the like).

The term “pharmaceutically acceptable salts” is meant to include salts of the active compounds that are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, e.g., Berge et al, Journal of Pharmaceutical Science 66: 1-19 (1977)). Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts. Other pharmaceutically acceptable carriers known to those of skill in the art are suitable for the present invention. Salts tend to be more soluble in aqueous or other protonic solvents that are the corresponding free base forms. In other cases, the preparation may be a lyophilized powder in a first buffer, e.g., in 1 mM-50 mM histidine, 0. 1%-2% sucrose, 2%-7% mannitol at a pH range of 4.5 to 5.5, that is combined with a second buffer prior to use.

Thus, the compounds of the present invention may exist as salts, such as with pharmaceutically acceptable acids. The present invention includes such salts. Examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, maleates, acetates, citrates, fumarates, tartrates (e.g., (+)-tartrates, (−)-tartrates, or mixtures thereof including racemic mixtures), succinates, benzoates, and salts with amino acids such as glutamic acid. These salts may be prepared by methods known to those skilled in the art.

The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.

The term “prodrug” may be used to describe compounds of the present invention that readily undergo chemical changes under physiological conditions to provide a biologically active compound that modulates eIF2B, eIF2α, or a component of the eIF2 or ISR pathways. Additionally, the prodrug compounds of the present invention can be converted to a biologically active compound that modulates eIF2B, eIF2α, or a component of the eIF2 or ISR pathways by chemical or biochemical methods in an ex vivo environment. For example, the prodrug compounds of the present invention may be slowly converted to a biologically active compound that modulates eIF2B, eIF2α, or a component of the eIF2 or ISR pathways when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

The term “pharmaceutically acceptable co-crystal” may be defined as a crystalline solid comprising two or more different compounds, wherein at least one of the two or more compounds is present in the crystalline solid in a neutral state (not ionized), wherein one of the two or more compounds is an active pharmaceutical ingredient (e.g., a compound of Formula (I)) and wherein the other compound or compounds present in the crystalline solid are also pharmaceutically acceptable. In some embodiments, at least two of the two or more compounds independently exist as solids under ambient conditions. In some embodiments, the two or more compounds interact through non-ionic intermolecular interactions.

Certain compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present invention.

Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

As used herein, the term “salt” refers to acid or base salts of the compounds used in the methods of the present invention. Illustrative examples of acceptable salts are mineral acid (hydrochloric acid, hydrobromic acid, phosphoric acid, and the like) salts, organic acid (acetic acid, propionic acid, glutamic acid, citric acid and the like) salts, quaternary ammonium (methyl iodide, ethyl iodide, and the like) salts.

Certain compounds of the present invention possess asymmetric carbon atoms (optical or chiral centers) or double bonds; the enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisometric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids, and individual isomers are encompassed within the scope of the present invention. The compounds of the present invention do not include those which are known in art to be too unstable to synthesize and/or isolate. The present invention is meant to include compounds in racemic and optically pure forms. Optically active (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain olefinic bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers.

As used herein, the term “isomers” refers to compounds having the same number and kind of atoms, and hence the same molecular weight, but differing in respect to the structural arrangement or configuration of the atoms.

The term “tautomer,” as used herein, refers to one of two or more structural isomers which exist in equilibrium and which are readily converted from one isomeric form to another.

It will be apparent to one skilled in the art that certain compounds of this invention may exist in tautomeric forms, all such tautomeric forms of the compounds being within the scope of the invention.

The terms “treating” or “treatment” refers to any indicia of success in the treatment or amelioration of an injury, disease, pathology or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; improving a patient's physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subjective parameters; including the results of a physical examination, neuropsychiatric exams, and/or a psychiatric evaluation. For example, certain methods herein treat cancer (e.g. pancreatic cancer, breast cancer, multiple myeloma, cancers of secretory cells), neurodegenerative diseases (e.g. Alzheimer's disease, Parkinson's disease, frontotemporal dementia), leukodystrophies (e.g., vanishing white matter disease, childhood ataxia with CNS hypo-myelination), postsurgical cognitive dysfunction, traumatic brain injury, hypoxia induced brain injury, stroke, spinal cord injury, intellectual disability syndromes, inflammatory diseases, musculoskeletal diseases, metabolic diseases, or diseases or disorders associated with impaired function of eIF2B or components in a signal transduction or signaling pathway including the ISR and decreased eIF2 pathway activity). For example certain methods herein treat cancer by decreasing or reducing or preventing the occurrence, growth, metastasis, or progression of cancer or decreasing a symptom of cancer; treat neurodegeneration by improving mental wellbeing, increasing mental function, slowing the decrease of mental function, decreasing dementia, delaying the onset of dementia, improving cognitive skills, decreasing the loss of cognitive skills, improving memory, decreasing the degradation of memory, decreasing a symptom of neurodegeneration or extending survival; treat vanishing white matter disease by reducing a symptom of vanishing white matter disease or reducing the loss of white matter or reducing the loss of myelin or increasing the amount of myelin or increasing the amount of white matter; treat childhood ataxia with CNS hypo-myelination by decreasing a symptom of childhood ataxia with CNS hypo-myelination or increasing the level of myelin or decreasing the loss of myelin; treat an intellectual disability syndrome by decreasing a symptom of an intellectual disability syndrome, treat an inflammatory disease by treating a symptom of the inflammatory disease; treat a musculoskeletal disease by treating a symptom of the musculoskeletal disease; treat a metabolic disease by treating a symptom of the metabolic disease; treat an autoimmune disease by treating a symptom of the autoimmune disease; treat an autoimmune disease by treating a symptom of the autoimmune disease; treat a viral infection by treating a symptom of the viral infection; treat a skin disease by treating a symptom of the skin disease; treat a fibrotic disease by treating a symptom of the fibrotic disease; treat a hemoglobin disease by treating a symptom of the hemoglobin disease; treat a hemoglobin disease by treating a symptom of the hemoglobin disease; treat a hearing loss condition by improving the hearing of a subject in need thereof; or treat an ocular disease by treating a symptom of the ocular disease or improving the vision of a subject in need thereof. Symptoms of a disease, disorder, or condition described herein (e.g., cancer a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, a metabolic disease, an autoimmune disease, a viral infection, a skin disease, a fibrotic disease, a hemoglobin disease, a kidney disease, a hearing loss condition, an ocular disease, or a condition or disease associated with impaired function of eIF2B or components in a signal transduction pathway including the eIF2 pathway, eIF2α phosphorylation, or ISR pathway) would be known or may be determined by a person of ordinary skill in the art. The term “treating” and conjugations thereof, include prevention of an injury, pathology, condition, or disease (e.g. preventing the development of one or more symptoms of a disease, disorder, or condition described herein).

An “effective amount” is an amount sufficient to accomplish a stated purpose (e.g. achieve the effect for which it is administered, treat a disease, reduce enzyme activity, increase enzyme activity, or reduce one or more symptoms of a disease or condition). An example of an “effective amount” is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease, which could also be referred to as a “therapeutically effective amount.” A “prophylactically effective amount” of a drug is an amount of a drug that, when administered to a subject, will have the intended prophylactic effect, e.g., preventing or delaying the onset (or reoccurrence) of an injury, disease, pathology or condition, or reducing the likelihood of the onset (or reoccurrence) of an injury, disease, pathology, or condition, or their symptoms. The full prophylactic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a prophylactically effective amount may be administered in one or more administrations. The exact amounts will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

A “reduction” of a symptom or symptoms (and grammatical equivalents of this phrase) means decreasing of the severity or frequency of the symptom(s), or elimination of the symptom(s).

The term “associated” or “associated with” in the context of a substance or substance activity or function associated with a disease (e.g., a disease or disorder described herein, e.g., cancer, a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, a metabolic disease, an autoimmune disease, a viral infection, a skin disease, a fibrotic disease, a hemoglobin disease, a kidney disease, a hearing loss condition, an ocular disease, or a disease or disorder associated with impaired function of eIF2B or components in a signal transduction pathway including the eIF2 pathway, eIF2a phosphorylation, or ISR pathway) means that the disease is caused by (in whole or in part), or a symptom of the disease is caused by (in whole or in part) the substance or substance activity or function. For example, a symptom of a disease or condition associated with an impaired function of the eIF2B may be a symptom that results (entirely or partially) from a decrease in eIF2B activity (e.g. decrease in eIF2B activity or levels, increase in eIF2α phosphorylation or activity of phosphorylated eIF2α or reduced eIF2 activity or increase in activity of phosphorylated eIF2a signal transduction or the ISR signaling pathway). As used herein, what is described as being associated with a disease, if a causative agent, could be a target for treatment of the disease. For example, a disease associated with decreased eIF2 activity or eIF2 pathway activity, may be treated with an agent (e.g., compound as described herein) effective for increasing the level or activity of eIF2 or eIF2 pathway or a decrease in phosphorylated eIF2α activity or the ISR pathway. For example, a disease associated with phosphorylated eIF2α may be treated with an agent (e.g., compound as described herein) effective for decreasing the level of activity of phosphorylated eIF2α or a downstream component or effector of phosphorylated eIF2α. For example, a disease associated with eIF2α, may be treated with an agent (e.g., compound as described herein) effective for increasing the level of activity of eIF2 or a downstream component or effector of eIF2.

“Control” or “control experiment” is used in accordance with its plain ordinary meaning and refers to an experiment in which the subjects or reagents of the experiment are treated as in a parallel experiment except for omission of a procedure, reagent, or variable of the experiment. In some instances, the control is used as a standard of comparison in evaluating experimental effects.

“Contacting” is used in accordance with its plain ordinary meaning and refers to the process of allowing at least two distinct species (e.g. chemical compounds including biomolecules, or cells) to become sufficiently proximal to react, interact or physically touch. It should be appreciated, however, that the resulting reaction product can be produced directly from a reaction between the added reagents or from an intermediate from one or more of the added reagents which can be produced in the reaction mixture. The term “contacting” may include allowing two species to react, interact, or physically touch, wherein the two species may be a compound as described herein and a protein or enzyme (e.g. eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway). In some embodiments contacting includes allowing a compound described herein to interact with a protein or enzyme that is involved in a signaling pathway (e.g. eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway).

As defined herein, the term “inhibition”, “inhibit”, “inhibiting” and the like in reference to a protein-inhibitor (e.g., antagonist) interaction means negatively affecting (e.g., decreasing) the activity or function of the protein relative to the activity or function of the protein in the absence of the inhibitor. In some embodiments, inhibition refers to reduction of a disease or symptoms of disease. In some embodiments, inhibition refers to a reduction in the activity of a signal transduction pathway or signaling pathway. Thus, inhibition includes, at least in part, partially or totally blocking stimulation, decreasing, preventing, or delaying activation, or inactivating, desensitizing, or down-regulating signal transduction or enzymatic activity or the amount of a protein. In some embodiments, inhibition refers to a decrease in the activity of a signal transduction pathway or signaling pathway (e.g., eIF2B, eIF2α, or a component of the eIF2 pathway, pathway activated by eIF2α phosphorylation, or ISR pathway). Thus, inhibition may include, at least in part, partially or totally decreasing stimulation, decreasing or reducing activation, or inactivating, desensitizing, or down-regulating signal transduction or enzymatic activity or the amount of a protein increased in a disease (e.g. eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway, wherein each is associated with cancer, a neurodegenerative disease, a leukodystrophy, an inflammatory disease, an autoimmune disease, a viral infection, a skin disease, a fibrotic disease, a hemoglobin disease, a kidney disease, a hearing loss condition, an ocular disease, a musculoskeletal disease, or a metabolic disease). Inhibition may include, at least in part, partially or totally decreasing stimulation, decreasing or reducing activation, or deactivating, desensitizing, or down-regulating signal transduction or enzymatic activity or the amount of a protein (e.g. eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway) that may modulate the level of another protein or increase cell survival (e.g., decrease in phosphorylated eIF2α pathway activity may increase cell survival in cells that may or may not have an increase in phosphorylated eIF2α pathway activity relative to a non-disease control or decrease in eIF2α pathway activity may increase cell survival in cells that may or may not have an increase in eIF2α pathway activity relative to a non-disease control).

As defined herein, the term “activation”, “activate”, “activating” and the like in reference to a protein-activator (e.g. agonist) interaction means positively affecting (e.g. increasing) the activity or function of the protein (e.g. eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway) relative to the activity or function of the protein in the absence of the activator (e.g. compound described herein). In some embodiments, activation refers to an increase in the activity of a signal transduction pathway or signaling pathway (e.g. eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway). Thus, activation may include, at least in part, partially or totally increasing stimulation, increasing or enabling activation, or activating, sensitizing, or up-regulating signal transduction or enzymatic activity or the amount of a protein decreased in a disease (e.g. level of eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway associated with cancer, a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, or a metabolic disease). Activation may include, at least in part, partially or totally increasing stimulation, increasing or enabling activation, or activating, sensitizing, or up-regulating signal transduction or enzymatic activity or the amount of a protein (e.g., eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway) that may modulate the level of another protein or increase cell survival (e.g., increase in eIF2α activity may increase cell survival in cells that may or may not have a reduction in eIF2α activity relative to a non-disease control).

The term “modulation” refers to an increase or decrease in the level of a target molecule or the function of a target molecule. In some embodiments, modulation of eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway may result in reduction of the severity of one or more symptoms of a disease associated with eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway (e.g., cancer, a neurodegenerative disease, a leukodystrophy, an inflammatory disease, an autoimmune disease, a viral infection, a skin disease, a fibrotic disease, a hemoglobin disease, a kidney disease, a hearing loss condition, an ocular disease, a musculoskeletal disease, or a metabolic disease) or a disease that is not caused by eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway but may benefit from modulation of eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway (e.g., decreasing in level or level of activity of eIF2B, eIF2a or a component of the eIF2 pathway).

The term “modulator” as used herein refers to modulation of (e.g., an increase or decrease in) the level of a target molecule or the function of a target molecule. In embodiments, a modulator of eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway is an anti-cancer agent. In embodiments, a modulator of eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway is a neuroprotectant. In embodiments, a modulator of eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway is a memory enhancing agent. In embodiments, a modulator of eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway is a memory enhancing agent (e.g., a long term memory enhancing agent). In embodiments, a modulator of eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway is an anti-inflammatory agent. In some embodiments, a modulator of eIF2B, eIF2α, or component of the eIF2 pathway or ISR pathway is a pain-relieving agent.

“Patient” or “subject” in need thereof refers to a living organism suffering from or prone to a disease or condition that can be treated by administration of a compound or pharmaceutical composition, as provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goat, sheep, cows, deer, and other non-mammalian animals. In some embodiments, a patient is human. In some embodiments, a patient is a domesticated animal. In some embodiments, a patient is a dog. In some embodiments, a patient is a parrot. In some embodiments, a patient is livestock animal. In some embodiments, a patient is a mammal. In some embodiments, a patient is a cat. In some embodiments, a patient is a horse. In some embodiments, a patient is bovine. In some embodiments, a patient is a canine. In some embodiments, a patient is a feline. In some embodiments, a patient is an ape. In some embodiments, a patient is a monkey. In some embodiments, a patient is a mouse. In some embodiments, a patient is an experimental animal. In some embodiments, a patient is a rat. In some embodiments, a patient is a hamster. In some embodiments, a patient is a test animal. In some embodiments, a patient is a newborn animal. In some embodiments, a patient is a newborn human. In some embodiments, a patient is a newborn mammal. In some embodiments, a patient is an elderly animal. In some embodiments, a patient is an elderly human. In some embodiments, a patient is an elderly mammal. In some embodiments, a patient is a geriatric patient.

“Disease”, “disorder” or “condition” refers to a state of being or health status of a patient or subject capable of being treated with a compound, pharmaceutical composition, or method provided herein. In some embodiments, the compounds and methods described herein comprise reduction or elimination of one or more symptoms of the disease, disorder, or condition, e.g., through administration of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

The term “signaling pathway” as used herein refers to a series of interactions between cellular and optionally extra-cellular components (e.g. proteins, nucleic acids, small molecules, ions, lipids) that conveys a change in one component to one or more other components, which in turn may convey a change to additional components, which is optionally propagated to other signaling pathway components.

“Pharmaceutically acceptable excipient” and “pharmaceutically acceptable carrier” refer to a substance that aids the administration of an active agent to and absorption by a subject and can be included in the compositions of the present invention without causing a significant adverse toxicological effect on the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with the compounds of the invention. One of skill in the art will recognize that other pharmaceutical excipients are useful in the present invention.

The term “preparation” is intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component with or without other carriers, is surrounded by a carrier, which is thus in association with it. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

As used herein, the term “administering” means oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intracranial, intranasal or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intra-arterial, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc. By “co-administer” it is meant that a composition described herein is administered at the same time, just prior to, or just after the administration of one or more additional therapies (e.g., anti-cancer agent, chemotherapeutic, or treatment for a neurodegenerative disease). The compound of the invention can be administered alone or can be coadministered to the patient. Coadministration is meant to include simultaneous or sequential administration of the compound individually or in combination (more than one compound or agent). Thus, the preparations can also be combined, when desired, with other active substances (e.g. to reduce metabolic degradation).

The term “eIF2B” as used herein refers to the heteropentameric eukaryotic translation initiation factor 2B. eIF2B is composed of five subunits: eIF2B1, eIF2B2, eIF2B3, eIF2B4 and eIF2B5. eIF2B1 refers to the protein associated with Entrez gene 1967, OMIM 606686, Uniprot Q14232, and/or RefSeq (protein) NP_001.405. eIF2B2 refers to the protein associated with Entrez gene 8892, OMIM 606454, Uniprot P49770, and/or RefSeq (protein) NP_055054. eIF2B3 refers to the protein associated with Entrez gene 8891, OMIM 606273, Uniprot Q9NR50, and/or RefSeq (protein) NP_065098. eIF2B4 refers to the protein associated with Entrez gene 8890, OMIM 606687, Uniprot Q9UI10, and/or RefSeq (protein) NP_751945. eIF2B5 refers to the protein associated with Entrez gene 8893, OMIM 603945, Uniprot Q13144, and/or RefSeq (protein) NP_003898.

The terms “eIF2alpha”, “eIF2a” or “eIF2α” are interchangeable and refer to the protein “eukaryotic translation initiation factor 2 alpha subunit eIF2S1”. In embodiments, “eIF2alpha”, “eIF2a” or “eIF2α” refer to the human protein. Included in the terms eIF2alpha”, “eIF2a” or “eIF2α” are the wildtype and mutant forms of the protein. In embodiments, “eIF2alpha”, “eIF2a” or “eIF2α” refer to the protein associated with Entrez Gene 1965, OMIM 603907, UniProt P05198, and/or RefSeq (protein) NP_004085. In embodiments, the reference numbers immediately above refer to the protein and associated nucleic acids known as of the date of filing of this application.

Compounds

In one aspect, provided herein is a compound represented by Formula (I):

or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, wherein:

R¹ is selected from the group consisting of —C(O)—C₁₋₄alkyl, —C(O)—O—C₁₋₄alkyl, —C(O)—N(R^(a))—C₁₋₄alkyl, —C(O)—C₁₋₄alkylene-C₁₋₄alkoxy, —C(O)—C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, -methylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—O—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—N(R^(a))—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-P(O)(OH)₂, —C(O)—C₁₋₅ alkylene-phenylene-O—P(O)(OH)₂, —C(O)—C₁₋₅ alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—C₁₋₅ alkylene-phenylene-(O—P(O)(OH)₂)(O—C₁₋₅ alkylene-P(O)(OH)₂), -methylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, -methylene-O—C(O)C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂. —C(O)—O—C₁₋₅alkylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, —C(O)—N(R^(a))-heteroarylene-C₁₋₂alkylene-O—P(O)(OH)₂, —P(O)(OH)₂, —SO₃H, —SO₂NR^(a)R^(b), —C(O)-heteroaryl, —C(O)—C₁₋₅alkylene-O—C₁₋₅alkylene-phenyl and methylene-C₁₋₅alkoxide;

wherein —C(O)—C₁₋₄alkyl is substituted by one or two substituents each independently selected from the group consisting of NR^(a)R^(b) and —CO₂H; and wherein —C(O)—O—C₁₋₄alkyl, —C(O)—N(R^(a))—C₁₋₄alkyl, —C(O)—C₁₋₄alkylene-C₁₋₄alkoxy, —C(O)—C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, methylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—O—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—N(R^(a))—C₁₋₅ alkylene-O—P(O)(OH)₂, —C(O)—C₁₋₅ alkyl ene-P (O)(OH)₂, —C(O)—C₁₋₅ alkylene-phenylene-O—P(O)(OH)₂, —C(O)—C₁₋₅ alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—C₁₋₅ alkylene-phenylene-(O—P(O)(OH)₂)(O—C₁₋₅ alkylene-P(O)(OH)₂), -methylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂ or -methylene-O—C(O)C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—O—C₁₋₅alkylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, —C(O)— heteroaryl, —C(O)—C₁₋₅alkylene-O—C₁₋₅alkylene-phenyl and —C(O)—N(R^(a))-heteroarylene-C₁₋₂alkylene-O—P(O)(OH)₂ may optionally be substituted by one, two, three or four substituents each independently selected from the group consisting of halogen, —CO₂H, —NR^(a)R^(b) and C₁₋₂alkyl (optionally substituted by one, two or three fluorines) and aryl; and

R^(a) and R^(b) are independently selected, for each occurrence, from the group consisting of hydrogen and C₁₋₃alkyl.

In some embodiments, R¹ is —C(O)—C₁₋₄alkyl; wherein —C(O)—C₁₋₄alkyl is substituted by one or two substituents each independently selected from the group consisting of NR^(a)R^(b) and —CO₂H.

In some embodiments, R¹ is selected from the group consisting of

In some embodiments, R¹ is —C(O)—O—C₁₋₄alkyl. In some embodiments, R¹ is selected from the group consisting of

In some embodiments, R¹ is —C(O)—N(R^(a))—C₁₋₄alkyl, wherein —C(O)—N(R^(a))—C₁₋₄alkyl may optionally be substituted by one or two CO₂H groups. In some embodiments, R¹ is represented by

In some embodiments, R¹ is —C(O)—C₁₋₄alkylene-C₁₋₄alkoxy. In some embodiments, R¹ is selected from the group consisting of

In some embodiments, R¹ is —C(O)—C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy. In some embodiments, R¹ is represented by

In some embodiments, R¹ is -methylene-O—P(O)(OH)₂. In some embodiments, R¹ is represented by

In some embodiments, R¹ is —C(O)—C₁₋₅alkylene-O—P(O)(OH)₂. In some embodiments, R¹ is selected from the group consisting of

In some embodiments, R¹ is —C(O)—O—C₁₋₅alkylene-O—P(O)(OH)₂. In some embodiments, R¹ is selected from the group consisting of

In some embodiments, R¹ is —C(O)—N(R^(a))—C₁₋₅alkylene-O—P(O)(OH)₂. In some embodiments, R¹ is selected from the group consisting of

In some embodiments, R¹ is —C(O)—C₁₋₅alkylene-P(O)(OH)₂. In some embodiments, R¹ is represented by

In some embodiments, R¹ is —C(O)—C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂ or —C(O)—C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)(O—C₁₋₅alkylene-P(O)(OH)₂). In some embodiments, R¹ is selected from the group consisting of

In some embodiments, R¹ is —C(O)—N(R^(a))-heteroarylene-C₁₋₂alkylene-O—P(O)(OH)₂. In some embodiments, R¹ is represented by

In some embodiments, R¹ is —C(O)-heteroaryl. In some embodiments, R¹ is selected from the group consisting of

In some embodiments, R¹ is —C(O)—C₁₋₅alkylene-O—C₁₋₅alkylene-phenyl. In some embodiments, R¹ is represented by

In some embodiments, R¹ is methylene-C₁₋₅alkoxide. In some embodiments, R¹ is represented by

In some embodiments, R¹ is —C(O)—O—C₁₋₅alkylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, -methylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂ or -methylene-O—C(O)C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂. In some embodiments, R¹ is represented by

In some embodiments, R¹ is selected from the group consisting of —P(O)(OH)₂, —SO₃H, and —SO₂NH₂.

In some embodiments, the compound of Formula (I) is selected from a compound set forth in Table 1 or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, or stereoisomer thereof.

TABLE 1 Exemplary compounds of the invention Compound Number Structure 100

101

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

134

135

136

137

138

139

140

141

Methods of Making Exemplary Compounds

The compounds of the present disclosure may be better understood in connection with the following synthetic schemes and methods which illustrate a means by which the compounds can be prepared. The compounds of the present disclosure can be prepared by a variety of synthetic procedures. Representative synthetic procedures are shown in, but not limited to, Schemes 1-5. The variables R¹, R^(a), and R^(b) are defined as detailed herein, e.g., in the Summary.

As shown in Scheme 1, compounds of formula (1-3), formula (1-5), and formula (1-7) can be prepared from compounds of formula (1-1).

Compounds of formula (1-1) can be reacted with a phosphoramidite of formula (1-2), wherein PG¹ is a protecting group such t-butyl or an optionally substituted benzyl, in the presence of an acetonitrile solution of 1H-tetrazole in an optionally heated solvent such as but not limited to N,N-dimethylformamide, dimethylacetamide or dichloromethane. Subsequent treatment with an oxidant such as hydrogen peroxide oxidizes the phosphorus moiety. Then the protecting groups, PG¹, can be removed using conditions known to one of skill in the art and dependent on the particular protecting group. For example, a benzyl protecting group can be removed by treatment by catalytic hydrogenation or treatment with an acid such as but not limited to trifluoroacetic acid give a compound of formula (1-3). For a t-butyl protecting group, treatment with trifluoroacetic acid in a solvent such dichloromethane also provides a compound of formula (1-3). A compound of formula (1-3) is representative of compounds of formula (I).

Compounds of formula (1-1) can be reacted with a C₁-3alkyl carbonochloridate of formula (1-4) in optionally warmed pyridine to give compounds of formula (1-5). Compounds of formula (1-5) are representative of compounds of formula (I).

Compounds of formula (1-1) can be reacted with compounds of formula (1-6), wherein R^(1-a) is OH or NR^(a)R^(b) in solvents such as dichloromethane or N,N-dimethylacetamide, respectively, to give compounds of formula (1-7). Compounds of formula (1-7) are representative of compounds of formula (I).

As shown in Scheme 2, compounds of formula (2-2) can be prepared from compounds of formula (1-1). Compounds of formula (1-1) can be coupled with carboxylic acids of formula (2-1) and subsequently deprotected if necessary to remove protective groups to give compounds of formula (2-2). Examples of conditions known to generate esters from a mixture of a carboxylic acid of formula (2-1) and an alcohol of formula (1-1) include but are not limited to adding a coupling reagent such as but not limited to 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, and bis(tetramethylene)fluoroformamidinium hexafluorophosphate. The coupling reagents may be added as a solid, a solution, or as the reagent bound to a solid support resin. In addition to the coupling reagents, auxiliary coupling reagents may facilitate the coupling reaction. Auxiliary coupling reagents that are often used in the coupling reactions include but are not limited to (dimethylamino)pyridine (DMAP). The reaction may be carried out optionally in the presence of a base such as triethylamine or N,N-diisopropylethylamine. The coupling reaction may be carried out in solvents such as but not limited to tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide, or dichloromethane. The coupling reaction can be carried out at ambient temperature or with heating, and the heating can be accomplished conventionally or with microwave irradiation. R^(2-a) is selected from the group consisting of —C₁₋₄alkyl, —C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₅alkylene-O—P(O)(OPG¹)₂, —C₁₋₅alkylene-P(O)(OPG¹)₂, —C₁₋₅alkylene-phenylene-O—P(O)(OPG¹)₂, and -heteroaryl; wherein —C₁₋₄alkyl is substituted by one or two substituents each independently selected from the group consisting of —NR^(a)PG² and —CO₂H; and wherein —C₁₋₄alkyl, —C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₅alkylene-O—P(O)(OPG¹)₂, —C₁₋₅alkylene-P(O)(OPG¹)₂, —C₁₋₅alkylene-phenylene-O—P(O)(OPG¹)₂, and heteroaryl may optionally be substituted by one, two, three or four substituents each independently selected from the group consisting of halogen, —CO₂H, C₁₋₂alkyl (optionally substituted by one, two or three fluorines) and aryl. The protecting group, PG¹, is as defined in Scheme 1. The protecting group, PG², is an amine protecting group such as but not limited to t-butoxycarbonyl. The protecting groups, PG¹ and PG², can be removed using conditions known to one of skill in the art and dependent on the particular protecting group. For example, a benzyl protecting group (PG) can be removed by treatment by catalytic hydrogenation or treatment with an acid such as but not limited to trifluoroacetic acid to give compounds of formula (2-2). For a t-butyl protecting group (PG) or t-butoxycarbonyl protecting group (PG²), treatment with trifluoroacetic acid in a solvent such dichloromethane also provides compounds of formula (2-2). Upon coupling or coupling followed by protecting group removal, compounds of formula (2-2) are obtained, wherein R^(2b) is —C₁₋₄alkyl, —C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₅alkylene-O—P(O)(OH)₂, —C₁₋₅alkylene-P(O)(OH)₂, —C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, and -heteroaryl; wherein —C₁₋₄alkyl is substituted by one or two substituents each independently selected from the group consisting of —NHR^(a) and —CO₂H; and wherein —C₁₋₄alkyl, —C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₄alkylene-O—C₁₋₄ alkylene-C₁₋₄alkoxy, —C₁₋₅ alkylene-O—P(O)(OH)₂, —C₁₋₅ alkylene-P(O)(OH)₂, —C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, and heteroaryl may optionally be substituted by one, two, three or four substituents each independently selected from the group consisting of halogen, —CO₂H, C₁₋₂alkyl (optionally substituted by one, two or three fluorines) and aryl. Compounds of formula (2-2) are representative of compounds of formula (I).

As shown in Scheme 3, compounds of formula (3-2) can be prepared from compounds of formula (1-1). Compounds of formula (1-1) can be coupled with carboxylic acid chlorides or chloroformates of formula (3-1) and subsequently deprotected if necessary to remove protective groups to give compounds of formula (3-2). The carboxylic acid chlorides can be obtained from commercial sources or prepared from the corresponding carboxylic acids by treatment with oxalyl chloride and a catalytic amount of N,N-dimethylformamide, thionyl chloride, cyanuric chloride, PCl₃ or PCl₅. Examples of conditions known to generate esters or carbonates from a mixture of a carboxylic acid chloride or chloroformates of formula (3-1) and an alcohol of formula (1-1) include but are not limited to adding a base such as triethylamine or N,N-diisopropylethylamine and an optional auxiliary coupling reagent such as but are not limited to (dimethylamino)pyridine (DMAP). The coupling reaction may be carried out in solvents such as but not limited to N,N-dimethylformamide, N,N-dimethylacetamide, and dichloromethane. R^(3-a) is selected from the group consisting of —C₁₋₄alkyl, —O—C₁₋₄alkyl, —C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₅alkylene-O—P(O)(OH)₂, —C₁₋₅alkylene-O—P(O)(OPG¹)₂, —O—C₁₋₅alkylene-O—P(O)(OPG¹)₂, —C₁₋₅alkylene-P(O)(OPG¹)₂, —C₁₋₅alkylene-phenylene-O—P(O)(OPG¹)₂, and -heteroaryl, wherein —C₁₋₄alkyl is substituted by one or two substituents each independently selected from the group consisting of NR^(a)PG² and —CO₂H; and wherein —O—C₁₋₄alkyl, —C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₅alkylene-O—P(O)(OH)₂, —C₁₋₅alkylene-O—P(O)(OPG¹)₂, —O—C₁₋₅alkylene-O—P(O)(OPG¹)₂, —C₁₋₅alkylene-P(O)(OPG¹)₂, —C₁₋₅alkylene-phenylene-O—P(O)(OPG¹)₂, and heteroaryl may optionally be substituted by one, two, three or four substituents each independently selected from the group consisting of halogen, —CO₂H, —NR^(a)R^(b), C₁₋₂alkyl (optionally substituted by one, two or three fluorines) and aryl. The protecting group, PG¹, is as defined in Scheme 1, and the protecting group, PG², is defined in Scheme 2. The protecting groups, PG¹ and PG², can be removed using conditions known to one of skill in the art and dependent on the particular protecting group as described in the previous schemes. Upon coupling or coupling followed by protecting group removal, compounds of formula (3-2) are obtained, wherein R^(3′) is —C₁₋₄alkyl, —O—C₁₋₄alkyl, —C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, —C₁₋₅alkylene-O—P(O)(OH)₂, —O—C₁₋₅alkylene-O—P(O)(OH)₂, —C₁₋₅alkylene-P(O)(OH)₂, —C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, and -heteroaryl; each optionally substituted as described above. In some instances, the esters or carbonates formed by the coupling may be further modified using methods known to one of skill in the art. Compounds of formula (3-2) are representative of compounds of formula (I).

As shown in Scheme 4, compounds of formula (4-2) can be prepared from compounds of formula (1-1). Compounds of formula (1-1) can be reacted with triphosgene or N,N′-disuccinimidyl carbonate optionally in pyridine or a mixture of pyridine and dichloromethane to give compounds of formula (4-1). Compounds of formula (4-1) can be reacted with nucleophiles, R^(4-b)—X¹—H (wherein X¹ is O or NR^(a) and R^(4-b) is —C₁₋₄alkyl, —C₁₋₅alkylene-O—P(O)(OPG¹)₂, -heteroarylene-C₁₋₂alkylene-O—P(O)(OPG¹)₂, wherein —C₁₋₄alkyl is substituted by one or two substituents each independently selected from the group consisting of bromine and —CO₂H), optionally in the presence of a base such as triethylamine or N,N-diisopropylethylamine or sodium bicarbonate and an optional auxiliary coupling reagent such as but are not limited to (dimethylamino)pyridine (DMAP) in a solvent such as tetrahydrofuran or ethyl acetate. The protecting group, PG¹, when present can be removed using conditions known to one of skill in the art and dependent on the particular protecting group as described in Scheme 1. In some instances, the carbonates or carbamates formed by the coupling may be further modified using methods known to one of skill in the art. For example, when R^(4-b) is an alkyl bromide, the bromine can be displaced in a nucleophilic substitution reaction. Compounds of formula (4-2) are representative of compounds of formula (I), wherein R^(4-c) is —C₁₋₄alkyl, —C₁₋₅alkylene-O—P(O)(OH)₂, -heteroarylene-C₁₋₂alkylene-O—P(O)(OH)₂, wherein —C₁₋₄alkyl is substituted by one or two —CO₂H. Compounds of formula (4-2) are representative of compounds of formula (I).

As shown in Scheme 5, compounds of formula (5-2) and formula (5-3) can be prepared from compounds of formula (1-1). Compounds of formula (1-1) can be reacted with chloromethyl methyl sulfide in the presence of iodine and a base such as sodium hydride in a solvent such as tetrahydrofuran to give compounds of formula (5-1). Compounds of formula (5-1) can be treated first with solid H₃PO₄ and activated 5 Å molecular sieves and then with N-iodosuccinimide to give a compound of formula (5-2). Alternatively, compounds of formula (5-1) can be treated first with solid H₃PO₄ and activated 5 Å molecular sieves and then with N-iodosuccinimide followed by a C₁₋₅alcohol to give compounds of formula (5-3). Compounds of formula (5-2) and formula (5-3) are representative of compounds of formula (I).

Pharmaceutical Compositions

The present invention features pharmaceutical compositions comprising a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is provided in an effective amount in the pharmaceutical composition. In some embodiments, the effective amount is a therapeutically effective amount. In certain embodiments, the effective amount is a prophylactically effective amount.

Pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. In general, such preparatory methods include the steps of bringing the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, (the “active ingredient”) into association with a carrier and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping and/or packaging the product into a desired single- or multi-dose unit. Pharmaceutical compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. As used herein, a “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.

Relative amounts of a compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered. By way of example, the composition may comprise between 0.1% and 100% (w/w) of a compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof.

The term “pharmaceutically acceptable excipient” refers to a non-toxic carrier, adjuvant, diluent, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable excipients useful in the manufacture of the pharmaceutical compositions of the invention are any of those that are well known in the art of pharmaceutical formulation and include inert diluents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Pharmaceutically acceptable excipients useful in the manufacture of the pharmaceutical compositions of the invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

Compositions of the present invention may be administered orally, parenterally (including subcutaneous, intramuscular, intravenous and intradermal), by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. In some embodiments, provided compounds or compositions are administrable intravenously and/or orally.

The term “parenteral” as used herein includes subcutaneous, intravenous, intramuscular, intraocular, intravitreal, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intraperitoneal intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, subcutaneously, intraperitoneally or intravenously. Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

Pharmaceutically acceptable compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added. In some embodiments, a provided oral formulation is formulated for immediate release or sustained/delayed release. In some embodiments, the composition is suitable for buccal or sublingual administration, including tablets, lozenges and pastilles. A compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may also be in micro-encapsulated form.

The compositions of the present invention can be delivered transdermally, by a topical route, formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols. Oral preparations include tablets, pills, powder, dragees, capsules, liquids, lozenges, cachets, gels, syrups, slurries, suspensions, etc., suitable for ingestion by the patient. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water/propylene glycol solutions. The compositions of the present invention may additionally include components to provide sustained release and/or comfort. Such components include high molecular weight, anionic mucomimetic polymers, gelling polysaccharides and finely-divided drug carrier substrates. These components are discussed in greater detail in U.S. Pat. Nos. 4,911,920; 5,403,841; 5,212, 162; and 4,861,760. The entire contents of these patents are incorporated herein by reference in their entirety for all purposes. The compositions of the present invention can also be delivered as microspheres for slow release in the body. For example, microspheres can be administered via intradermal injection of drug-containing microspheres, which slowly release subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645, 1995; as biodegradable and injectable gel formulations (see, e.g., Gao Pharm. Res. 12:857-863, 1995); or, as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674, 1997). In another embodiment, the formulations of the compositions of the present invention can be delivered by the use of liposomes which fuse with the cellular membrane or are endocytosed, i.e., by employing receptor ligands attached to the liposome, that bind to surface membrane protein receptors of the cell resulting in endocytosis. By using liposomes, particularly where the liposome surface carries receptor ligands specific for target cells, or are otherwise preferentially directed to a specific organ, one can focus the delivery of the compositions of the present invention into the target cells in vivo. (See, e.g., Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, J. Hosp. Pharm. 46: 1576-1587, 1989). The compositions of the present invention can also be delivered as nanoparticles.

Alternatively, pharmaceutically acceptable compositions of this invention may be administered in the form of suppositories for rectal administration. Pharmaceutically acceptable compositions of this invention may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

In some embodiments, in order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with ordinary experimentation.

Compounds provided herein, e.g., a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof are typically formulated in dosage unit form, e.g., single unit dosage form, for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject or organism will depend upon a variety of factors including the disease being treated and the severity of the disorder; the activity of the specific active ingredient employed; the specific composition employed; the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of the treatment; drugs used in combination or coincidental with the specific active ingredient employed; and like factors well known in the medical arts.

The exact amount of a compound required to achieve an effective amount will vary from subject to subject, depending, for example, on species, age, and general condition of a subject, severity of the side effects or disorder, identity of the particular compound(s), mode of administration, and the like. The desired dosage can be delivered three times a day, two times a day, once a day, every other day, every third day, every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the desired dosage can be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations).

In certain embodiments, an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof for administration one or more times a day may comprise about 0.0001 mg to about 5000 mg, e.g., from about 0.0001 mg to about 4000 mg, about 0.0001 mg to about 2000 mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg, about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1 mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about 1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosage form.

In certain embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be at dosage levels sufficient to deliver from about 0.001 mg/kg to about 1000 mg/kg, e.g., about 0.001 mg/kg to about 500 mg/kg, about 0.01 mg/kg to about 250 mg/kg, about 0.1 mg/kg to about 100 mg/kg, about 0.1 mg/kg to about 50 mg/kg, about 0.1 mg/kg to about 40 mg/kg, about 0.1 mg/kg to about 25 mg/kg, about 0.01 mg/kg to about 10 mg/kg, about 0.1 mg/kg to about 10 mg/kg, or about 1 mg/kg to about 50 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.

It will be appreciated that dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult. The amount to be administered to, for example, a child or an adolescent can be determined by a medical practitioner or person skilled in the art and can be lower or the same as that administered to an adult.

It will be also appreciated that a compound or composition, e.g., a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof as described herein, can be administered in combination with one or more additional pharmaceutical agents. The compounds or compositions can be administered in combination with additional pharmaceutical agents that improve their bioavailability, reduce and/or modify their metabolism, inhibit their excretion, and/or modify their distribution within the body. It will also be appreciated that the therapy employed may achieve a desired effect for the same disorder, and/or it may achieve different effects.

The compound or composition can be administered concurrently with, prior to, or subsequent to, one or more additional pharmaceutical agents, which may be useful as, e.g., combination therapies. Pharmaceutical agents include therapeutically active agents. Pharmaceutical agents also include prophylactically active agents. Each additional pharmaceutical agent may be administered at a dose and/or on a time schedule determined for that pharmaceutical agent. The additional pharmaceutical agents may also be administered together with each other and/or with the compound or composition described herein in a single dose or administered separately in different doses. The particular combination to employ in a regimen will take into account compatibility of the inventive compound with the additional pharmaceutical agents and/or the desired therapeutic and/or prophylactic effect to be achieved. In general, it is expected that the additional pharmaceutical agents utilized in combination be utilized at levels that do not exceed the levels at which they are utilized individually. In some embodiments, the levels utilized in combination will be lower than those utilized individually.

Exemplary additional pharmaceutical agents include, but are not limited to, anti-proliferative agents, anti-cancer agents, anti-diabetic agents, anti-inflammatory agents, immunosuppressant agents, and pain-relieving agents. Pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved by the U.S. Food and Drug Administration as provided in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells.

Pharmaceutical compositions provided by the present invention include compositions wherein the active ingredient (e.g., compounds described herein, including embodiments or examples) is contained in a therapeutically effective amount, i.e., in an amount effective to achieve its intended purpose. The actual amount effective for a particular application will depend, inter alia, on the condition being treated. When administered in methods to treat a disease, such compositions will contain an amount of active ingredient effective to achieve the desired result, e.g., modulating the activity of a target molecule (e.g. eIF2B, eIF2 or component of eIF2α signal transduction pathway or component of phosphorylated eIF2α pathway or the ISR pathway), and/or reducing, eliminating, or slowing the progression of disease symptoms (e.g. symptoms of cancer a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, a metabolic disease, or a disease or disorder associated with impaired function of eIF2B, eIF2α or a component of the eIF2 pathway or ISR pathway). Determination of a therapeutically effective amount of a compound of the invention is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure herein.

The dosage and frequency (single or multiple doses) administered to a mammal can vary depending upon a variety of factors, for example, whether the mammal suffers from another disease, and its route of administration; size, age, sex, health, body weight, body mass index, and diet of the recipient; nature and extent of symptoms of the disease being treated (e.g. a symptom of cancer, a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, a metabolic disease, or a disease or disorder associated with impaired function of eIF2B, eIF2 α, or a component of the eIF2 pathway or ISR pathway), kind of concurrent treatment, complications from the disease being treated or other health-related problems. Other therapeutic regimens or agents can be used in conjunction with the methods and compounds of Applicants' invention. Adjustment and manipulation of established dosages (e.g., frequency and duration) are well within the ability of those skilled in the art.

For any compound described herein, the therapeutically effective amount can be initially determined from cell culture assays. Target concentrations will be those concentrations of active compound(s) that are capable of achieving the methods described herein, as measured using the methods described herein or known in the art.

As is well known in the art, therapeutically effective amounts for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals. The dosage in humans can be adjusted by monitoring compounds effectiveness and adjusting the dosage upwards or downwards, as described above. Adjusting the dose to achieve maximal efficacy in humans based on the methods described above and other methods is well within the capabilities of the ordinarily skilled artisan.

Dosages may be varied depending upon the requirements of the patient and the compound being employed. The dose administered to a patient, in the context of the present invention should be sufficient to affect a beneficial therapeutic response in the patient over time. The size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects. Determination of the proper dosage for a particular situation is within the skill of the practitioner. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under circumstances is reached. Dosage amounts and intervals can be adjusted individually to provide levels of the administered compound effective for the particular clinical indication being treated. This will provide a therapeutic regimen that is commensurate with the severity of the individual's disease state.

Utilizing the teachings provided herein, an effective prophylactic or therapeutic treatment regimen can be planned that does not cause substantial toxicity and yet is effective to treat the clinical symptoms demonstrated by the particular patient. This planning should involve the careful choice of active compound by considering factors such as compound potency, relative bioavailability, patient body weight, presence and severity of adverse side effects, preferred mode of administration and the toxicity profile of the selected agent.

Also encompassed by the invention are kits (e.g., pharmaceutical packs). The inventive kits may be useful for preventing and/or treating a disease (e.g., cancer, a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, a metabolic disease, or other disease or condition described herein).

The kits provided may comprise an inventive pharmaceutical composition or compound and a container (e.g., a vial, ampule, bottle, syringe, and/or dispenser package, or other suitable container). In some embodiments, provided kits may optionally further include a second container comprising a pharmaceutical excipient for dilution or suspension of an inventive pharmaceutical composition or compound. In some embodiments, the inventive pharmaceutical composition or compound provided in the container and the second container are combined to form one unit dosage form.

Thus, in one aspect, provided are kits including a first container comprising a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, or a pharmaceutical composition thereof. In certain embodiments, the kits are useful in preventing and/or treating a proliferative disease in a subject. In certain embodiments, the kits further include instructions for administering a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, or a pharmaceutical composition thereof, to a subject to prevent and/or treat a disease described herein.

Methods of Treatment

The present invention features compounds, compositions, and methods comprising a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof. In some embodiments, the compounds, compositions, and methods are used in the prevention or treatment of a disease, disorder, or condition. Exemplary diseases, disorders, or conditions include, but are not limited to a neurodegenerative disease, a leukodystrophy, a cancer, an inflammatory disease, an autoimmune disease, a viral infection, a skin disease, a fibrotic disease, a hemoglobin disease, a kidney disease, a hearing loss condition, an ocular disease, a disease with mutations that leads to UPR induction, a malaria infection, a musculoskeletal disease, a metabolic disease, or a mitochondrial disease.

In some embodiments, the disease, disorder, or condition is related to (e.g., caused by) modulation of (e.g., a decrease in) eIF2B activity or level, eIF2α activity or level, or a component of the eIF2 pathway or ISR pathway. In some embodiments, the disease, disorder, or condition is related to modulation of a signaling pathway related to a component of the eIF2 pathway or ISR pathway (e.g., phosphorylation of a component of the eIF2 pathway or ISR pathway). In some embodiments, the disease, disorder, or condition is related to (e.g., caused by) neurodegeneration. In some embodiments, the disease, disorder, or condition is related to (e.g., caused by) neural cell death or dysfunction. In some embodiments, the disease, disorder, or condition is related to (e.g., caused by) glial cell death or dysfunction. In some embodiments, the disease, disorder, or condition is related to (e.g., caused by) an increase in the level or activity of eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway. In some embodiments, the disease, disorder, or condition is related to (e.g., caused by) a decrease in the level or activity of eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway.

In some embodiments, the disease may be caused by a mutation to a gene or protein sequence related to a member of the eIF2 pathway (e.g., eIF2B, eIF2α, or other component). Exemplary mutations include an amino acid mutation in the eIF2B1, eIF2B2, eIF2B3, eIF2B4, eIF2B5 subunits. In some embodiments, an amino acid mutation (e.g., an amino acid substitution, addition, or deletion) in a particular protein that may result in a structural change, e.g., a conformational or steric change, that affects the function of the protein. For example, in some embodiments, amino acids in and around the active site or close to a binding site (e.g., a phosphorylation site, small molecule binding site, or protein-binding site) may be mutated such that the activity of the protein is impacted. In some instances, the amino acid mutation (e.g., an amino acid substitution, addition, or deletion) may be conservative and may not substantially impact the structure or function of a protein. For example, in certain cases, the substitution of a serine residue with a threonine residue may not significantly impact the function of a protein. In other cases, the amino acid mutation may be more dramatic, such as the substitution of a charged amino acid (e.g., aspartic acid or lysine) with a large, nonpolar amino acid (e.g., phenylalanine or tryptophan) and therefore may have a substantial impact on protein function. The nature of the mutations that affect the structure of function of a gene or protein may be readily identified using standard sequencing techniques, e.g., deep sequencing techniques that are well known in the art. In some embodiments, a mutation in a member of the eIF2 pathway may affect binding or activity of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof and thereby modulate treatment of a particular disease, disorder, or condition, or a symptom thereof.

In some embodiments, an eIF2 protein may comprise an amino acid mutation (e.g., an amino acid substitution, addition, or deletion) at an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine residue. In some embodiments, an eIF2 protein may comprise an amino acid substitution at an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine residue. In some embodiments, an eIF2 protein may comprise an amino acid addition at an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine residue. In some embodiments, an eIF2 protein may comprise an amino acid deletion at an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine residue.

In some embodiments, the eIF2 protein may comprise an amino acid mutation (e.g., an amino acid substitution, addition, or deletion) at an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine residue in the eIF2B1, eIF2B2, eIF2B3, eIF2B4, eIF2B5 subunits. In some embodiments, the eIF2 protein may comprise an amino acid substitution at an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine residue in the eIF2B1, eIF2B2, eIF2B3, eIF2B4, eIF2B5 subunits. In some embodiments, the eIF2 protein may comprise an amino acid addition at an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine residue in the eIF2B1, eIF2B2, eIF2B3, eIF2B4, eIF2B5 subunits. In some embodiments, the eIF2 protein may comprise an amino acid deletion at an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine residue in the eIF2B1, eIF2B2, eIF2B3, eIF2B4, eIF2B5 subunits. Exemplary mutations include V183F (eIF2B1 subunit), H341Q (eIF2B3), I346T (eIF2B3), R483W (eIF2B4), R113H (eIF2B5), and R195H (eIF2B5).

In some embodiments, an amino acid mutation (e.g., an amino acid substitution, addition, or deletion) in a member of the eIF2 pathway (e.g., an eIF2B protein subunit) may affect binding or activity of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof and thereby modulate treatment of a particular disease, disorder, or condition, or a symptom thereof.

Neurodegenerative Disease

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a neurodegenerative disease. As used herein, the term “neurodegenerative disease” refers to a disease or condition in which the function of a subject's nervous system becomes impaired. Examples of a neurodegenerative disease that may be treated with a compound, pharmaceutical composition, or method described herein include Alexander's disease, Alper's disease, Alzheimer's disease, Amyotrophic lateral sclerosis (ALS), Ataxia telangiectasia, Batten disease (also known as Spielmeyer-Vogt-Sjogren-Batten disease), Bovine spongiform encephalopathy (BSE), Canavan disease, Cockayne syndrome, Corticobasal degeneration, Creutzfeldt-Jakob disease, Dystonia, frontotemporal dementia (FTD), Gerstmann-Straussler-Scheinker syndrome, Huntington's disease, HIV-associated dementia, Kennedy's disease, Krabbe disease, kuru, Lewy body dementia, Machado-Joseph disease (Spinocerebellar ataxia type 3), Multiple system atrophy, Multisystem proteinopathy, Narcolepsy, Neuroborreliosis, Parkinson's disease, Pelizaeus-Merzbacher Disease, Pick's disease, Primary lateral sclerosis, Prion diseases, Refsum's disease, Sandhoff disease, Schilder's disease, Subacute combined degeneration of spinal cord secondary to Pernicious Anaemia, Schizophrenia, Spinocerebellar ataxia (multiple types with varying characteristics, e.g., Spinocerebellar ataxia type 2 or Spinocerebellar ataxia type 8), Spinal muscular atrophy, Steele-Richardson-Olszewski disease, progressive supranuclear palsy, corticobasal degeneration, adrenoleukodystrophy, X-linked adrenoleukodystrophy, cerebral adrenoleukodystrophy, Pelizaeus-Merzbacher Disease, Krabbe disease, leukodystrophy due to mutation in DARS2 gene (sometimes known as lukoencephalopathy with brainstem and spinal cord involvement and lactate elevation (LBSL), DARS2-related spectrum disorders, or Tabes dorsalis.

In some embodiments, the neurodegenerative disease comprises vanishing white matter disease, childhood ataxia with CNS hypo-myelination, a leukodystrophy, a leukoencephalopathy, a hypomyelinating or demyelinating disease, an intellectual disability syndrome (e.g., Fragile X syndrome), Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Creutzfeldt-Jakob disease, frontotemporal dementia (FTD), Gerstmann-Straussler-Scheinker disease, Huntington's disease, dementia (e.g., HIV-associated dementia or Lewy body dementia), kuru, multiple sclerosis, Parkinson's disease, or a prion disease.

In some embodiments, the neurodegenerative disease comprises vanishing white matter disease, childhood ataxia with CNS hypo-myelination, a leukodystrophy, a leukoencephalopathy, a hypomyelinating or demyelinating disease, or an intellectual disability syndrome (e.g., Fragile X syndrome).

In some embodiments, the neurodegenerative disease comprises a psychiatric disease such as agoraphobia, Alzheimer's disease, anorexia nervosa, amnesia, anxiety disorder, attention deficit disorder, bipolar disorder, body dysmorphic disorder, bulimia nervosa, claustrophobia, depression, delusions, Diogenes syndrome, dyspraxia, insomnia, Munchausen's syndrome, narcolepsy, narcissistic personality disorder, obsessive-compulsive disorder, psychosis, phobic disorder, schizophrenia, seasonal affective disorder, schizoid personality disorder, sleepwalking, social phobia, substance abuse, tardive dyskinesia, Tourette syndrome, or trichotillomania.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat vanishing white matter disease. Exemplary methods of treating vanishing white matter disease include, but are not limited to, reducing or eliminating a symptom of vanishing white matter disease, reducing the loss of white matter, reducing the loss of myelin, increasing the amount of myelin, or increasing the amount of white matter in a subject.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat childhood ataxia with CNS hypo-myelination. Exemplary methods of treating childhood ataxia with CNS hypo-myelination include, but are not limited to, reducing or eliminating a symptom of childhood ataxia with CNS hypo-myelination, increasing the level of myelin, or decreasing the loss of myelin in a subject.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat an intellectual disability syndrome (e.g., Fragile X syndrome). Exemplary methods of treating an intellectual disability syndrome include, but are not limited to, reducing or eliminating a symptom of an intellectual disability syndrome.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat neurodegeneration. Exemplary methods of treating neurodegeneration include, but are not limited to, improvement of mental wellbeing, increasing mental function, slowing the decrease of mental function, decreasing dementia, delaying the onset of dementia, improving cognitive skills, decreasing the loss of cognitive skills, improving memory, decreasing the degradation of memory, or extending survival.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a leukoencephalopathy or demyelinating disease. Exemplary leukoencephalopathies include, but are not limited to, progressive multifocal leukoencephalopathy, toxic leukoencephalopathy, leukoencephalopathy with vanishing white matter, leukoencephalopathy with neuroaxonal spheroids, reversible posterior leukoencephalopathy syndrome, hypertensive leukoencephalopathy, megalencephalic leukoencephalopathy with subcortical cysts, Charcot-Marie-Tooth disorder, and Devic's disease. A leukoencephalopathy may comprise a demyelinating disease, which may be inherited or acquired. In some embodiments, an acquired demyelinating disease may be an inflammatory demyelinating disease (e.g., an infectious inflammatory demyelinating disease or a non-infectious inflammatory demyelinating disease), a toxic demyelinating disease, a metabolic demyelinating disease, a hypoxic demyelinating disease, a traumatic demyelinating disease, or an ischemic demyelinating disease (e.g., Binswanger's disease). Exemplary methods of treating a leukoencephalopathy or demyelinating disease include, but are not limited to, reducing or eliminating a symptom of a leukoencephalopathy or demyelinating disease, reducing the loss of myelin, increasing the amount of myelin, reducing the loss of white matter in a subject, or increasing the amount of white matter in a subject.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a traumatic injury or a toxin-induced injury to the nervous system (e.g., the brain). Exemplary traumatic brain injuries include, but are not limited to, a brain abscess, concussion, ischemia, brain bleeding, cranial fracture, diffuse axonal injury, locked-in syndrome, or injury relating to a traumatic force or blow to the nervous system or brain that causes damage to an organ or tissue. Exemplary toxin-induced brain injuries include, but are not limited to, toxic encephalopathy, meningitis (e.g. bacterial meningitis or viral meningitis), meningoencephalitis, encephalitis (e.g., Japanese encephalitis, eastern equine encephalitis, West Nile encephalitis), Guillan-Barre syndrome, Sydenham's chorea, rabies, leprosy, neurosyphilis, a prion disease, or exposure to a chemical (e.g., arsenic, lead, toluene, ethanol, manganese, fluoride, dichlorodiphenyltrichloroethane (DDT), dichlorodiphenyldichloroethylene (DDE), tetrachloroethylene, a polybrominated diphenyl ether, a pesticide, a sodium channel inhibitor, a potassium channel inhibitor, a chloride channel inhibitor, a calcium channel inhibitor, or a blood brain barrier inhibitor).

In other embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to improve memory in a subject. Induction of memory has been shown to be facilitated by decreased and impaired by increased eIF2α phosphorylation. Regulators of translation, such as compounds disclosed herein (e.g. a compound of Formula (I)), could serve as therapeutic agents that improve memory in human disorders associated with memory loss such as Alzheimer's disease and in other neurological disorders that activate the UPR or ISR in neurons and thus could have negative effects on memory consolidation such as Parkinson's disease, schizophrenia, amyotrophic lateral sclerosis (ALS) and prion diseases. In addition, a mutation in eIF2γ that disrupts complex integrity linked intellectual disability (intellectual disability syndrome or ID) to impaired translation initiation in humans. Hence, two diseases with impaired eIF2 function, ID and VWM, display distinct phenotypes but both affect mainly the brain and impair learning. In some embodiments, the disease or condition is unsatisfactory memory (e.g., working memory, long term memory, short term memory, or memory consolidation).

In still other embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used in a method to improve memory in a subject (e.g., working memory, long term memory, short term memory, or memory consolidation). In some embodiments, the subject is human. In some embodiments, the subject is a non-human mammal. In some embodiments, the subject is a domesticated animal. In some embodiments, the subject is a dog. In some embodiments, the subject is a bird. In some embodiments, the subject is a horse. In embodiments, the patient is a bovine. In some embodiments, the subject is a primate.

Cancer

In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I), Formula (II) or Formula (III) is used to treat cancer. As used herein, “cancer” refers to human cancers and carcinomas, sarcomas, adenocarcinomas (e.g., papillary adenocarcinomas), lymphomas, leukemias, melanomas, etc., including solid and lymphoid cancers, kidney, breast, lung, bladder, colon, ovarian, prostate, pancreas, stomach, brain, head and neck, skin, uterine, testicular, glioma, esophagus, liver cancer, including hepatocarcinoma, lymphoma, including B-acute lymphoblastic lymphoma, non-Hodgkin's lymphomas (e.g., Burkitt's, Small Cell, and Large Cell lymphomas), Hodgkin's lymphoma, leukemia (including AML, ALL, and CML), and/or multiple myeloma. In some further instances, “cancer” refers to lung cancer, breast cancer, ovarian cancer, epithelial ovarian cancer, leukemia, lymphoma, melanoma, pancreatic cancer, sarcoma, bladder cancer, bone cancer, biliary tract cancer, adrenal gland cancer, salivary gland cancer, bronchus cancer, oral cancer, cancer of the oral cavity or pharynx, laryngeal cancer, renal cancer, gynecologic cancers, brain cancer, central nervous system cancer, peripheral nervous system cancer, cancer of the hematological tissues, small bowel or appendix cancer, cervical cancer, colon cancer, esophageal cancer, gastric cancer, liver cancer, head and neck cancer, kidney cancer, myeloma, thyroid cancer, prostate cancer, metastatic cancer, or carcinoma.

As used herein, the term “cancer” refers to all types of cancer, neoplasm or malignant tumors found in mammals, including leukemia, lymphoma, carcinomas and sarcomas. Exemplary cancers that may be treated with a compound, pharmaceutical composition, or method provided herein include lymphoma, B-cell lymphoma, heavy chain disease, alpha chain disease, gamma chain disease, mu chain disease, Waldenstrom's macroglobulinemia, benign monoclonal gammopathy, sarcoma, bladder cancer, bone cancer, brain tumor, cervical cancer, colon cancer, esophageal cancer, gastric cancer, head and neck cancer, kidney cancer, myeloma, thyroid cancer, leukemia, prostate cancer, breast cancer (e.g., ER positive, ER negative, chemotherapy resistant, herceptin resistant, HER2 positive, doxorubicin resistant, tamoxifen resistant, ductal carcinoma, lobular carcinoma, primary, metastatic), ovarian cancer, pancreatic cancer, liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., non-small cell lung carcinoma, squamous cell lung carcinoma, adenocarcinoma, large cell lung carcinoma, small cell lung carcinoma, carcinoid, sarcoma), glioblastoma multiforme, acoustic neuroma, retinoblastoma, astrocytoma, craniopharyngioma, hemangioblastoma, pinealoma, ependymoma, oligodendroglioma, meningioma, glioma, or melanoma. Additional examples include, cancer of the thyroid, endocrine system, brain, breast, cervix, colon, head & neck, liver, kidney, lung, non-small cell lung, melanoma, mesothelioma, ovary, sarcoma, stomach, uterus or Medulloblastoma, Hodgkin's Disease, Non-Hodgkin's Lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, immunocytic amyloidosis, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, primary brain tumors, cancer, malignant pancreatic insulanoma, malignant carcinoid, urinary bladder cancer, premalignant skin lesions, testicular cancer, lymphomas, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, endometrial cancer, adrenal cortical cancer, neoplasms of the endocrine or exocrine pancreas, medullary thyroid cancer, medullary thyroid carcinoma, melanoma, colorectal cancer, papillary thyroid cancer, hepatocellular carcinoma, Paget's Disease of the Nipple, Phyllodes Tumors, Lobular Carcinoma, Ductal Carcinoma, cancer of the pancreatic stellate cells, cancer of the hepatic stellate cells, or prostate cancer.

The term “leukemia” refers broadly to progressive, malignant diseases of the blood-forming organs and is generally characterized by a distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is generally clinically classified on the basis of (1) the duration and character of the disease-acute or chronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid (lymphogenous), or monocytic; and (3) the increase or non-increase in the number abnormal cells in the blood-leukemic or aleukemic (subleukemic). Exemplary leukemias that may be treated with a compound, pharmaceutical composition, or method provided herein include, for example, chronic leukemia, acute nonlymphocytic leukemia, acute lymphocytic leukemia, B-cell chronic lympliocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, a leukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovine leukemia, acute myelocytic leukemia, chronic myelocytic leukemia, leukemia cutis, embryonal leukemia, eosinophilic leukemia, erythroleukemia, Gross' leukemia, hairy-cell leukemia, hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphogenous leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocyte leukemia, micromyeloblastic leukemia, monocytic leukemia, myeloblasts leukemia, myelocytic leukemia, myeloid granulocytic leukemia, myelomonocytic leukemia, Naegeli leukemia, plasma cell leukemia, multiple myeloma, plasmacytic leukemia, polycythemia vera, promyelocytic leukemia, Rieder cell leukemia, Schilling's leukemia, stem cell leukemia, subleukemic leukemia, or undifferentiated cell leukemia.

The term “sarcoma” generally refers to a tumor which is made up of a substance like the embryonic connective tissue and is generally composed of closely packed cells embedded in a fibrillar or homogeneous substance. Sarcomas that may be treated with a compound, pharmaceutical composition, or method provided herein include a chondrosarcoma, fibrosarcoma, leiomyosarcoma, lymphosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Abemethy's sarcoma, adipose sarcoma, liposarcoma, alveolar soft part sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma, chorio carcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrial sarcoma, endotheliosarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblastic sarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, immunoblastic sarcoma of B cells, lymphoma, immunoblastic sarcoma of T-cells, Jensen's sarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukosarcoma, malignant mesenchymoma sarcoma, osteogenic sarcoma, parosteal sarcoma, reticulocytic sarcoma, Rous sarcoma, serocystic sarcoma, synovial sarcoma, or telangiectaltic sarcoma.

The term “melanoma” is taken to mean a tumor arising from the melanocytic system of the skin and other organs. Melanomas that may be treated with a compound, pharmaceutical composition, or method provided herein include, for example, acral-lentiginous melanoma, amelanotic melanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma, Harding-Passey melanoma, juvenile melanoma, lentigo maligna melanoma, malignant melanoma, nodular melanoma, subungal melanoma, or superficial spreading melanoma.

The term “carcinoma” refers to a malignant new growth made up of epithelial cells tending to infiltrate the surrounding tissues and give rise to metastases. Exemplary carcinomas that may be treated with a compound, pharmaceutical composition, or method provided herein include, for example, medullary thyroid carcinoma, familial medullary thyroid carcinoma, acinar carcinoma, acinous carcinoma, adenocystic carcinoma, adenoid cystic carcinoma, carcinoma adenomatosum, carcinoma of adrenal cortex, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, carcinoma basocellulare, basaloid carcinoma, basosquamous cell carcinoma, bile duct carcinoma, bladder carcinoma, breast carcinoma, Brenner carcinoma, bronchioalveolar carcinoma, bronchiolar carcinoma, bronchiogenic carcinoma, cerebriform carcinoma, cervical carcinoma, cholangiocellular carcinoma, chordoma, chorionic carcinoma, clear cell carcinoma, colloid carcinoma, colon carcinoma, comedo carcinoma, corpus carcinoma, cribriform carcinoma, carcinoma en cuirasse, carcinoma cutaneum, cylindrical carcinoma, cylindrical cell carcinoma, cystadenocarcinoma, duct carcinoma, ductal carcinoma, carcinoma durum, embryonal carcinoma, encephaloid carcinoma, endometrioid carcinoma, epiermoid carcinoma, epithelial carcinoma, carcinoma epitheliale adenoides, exophytic carcinoma, carcinoma ex ulcere, carcinoma fibrosum, gelatiniforni carcinoma, gelatinous carcinoma, giant cell carcinoma, carcinoma gigantocellulare, glandular carcinoma, granulosa cell carcinoma, hair-matrix carcinoma, hematoid carcinoma, hepatoma, hepatocellular carcinoma, Hurthle cell carcinoma, hyaline carcinoma, hypernephroid carcinoma, infantile embryonal carcinoma, carcinoma in situ, intraepidermal carcinoma, intraepithelial carcinoma, Krompecher's carcinoma, Kulchitzky-cell carcinoma, large-cell carcinoma, lenticular carcinoma, carcinoma lenticulare, lipomatous carcinoma, lobular carcinoma, lung carcinoma, lymphoepithelial carcinoma, carcinoma medullare, medullary carcinoma, melanotic carcinoma, carcinoma molle, mucinous carcinoma, carcinoma muciparum, carcinoma mucocellulare, mucoepidermoid carcinoma, carcinoma mucosum, mucous carcinoma, carcinoma myxomatodes, nasopharyngeal carcinoma, nonpapillary renal cell carcinoma, oat cell carcinoma, carcinoma ossificans, osteoid carcinoma, ovarian carcinoma, pancreatic ductal caiciiioiria, papillary carcinoma, periportal carcinoma, preinvasive carcinoma, prickle cell carcinoma, pultaceous carcinoma, renal cell carcinoma of kidney, reserve cell carcinoma, carcinoma sarcomatodes, schneiderian carcinoma, scirrhous carcinoma, carcinoma scroti, sebaceous gland carcinoma, seminoma, serous carcinoma, signet-ring cell carcinoma, carcinoma simplex, small-cell carcinoma, solanoid carcinoma, spheroidal cell carcinoma, spindle cell carcinoma, carcinoma spongiosum, squamous carcinoma, squamous cell carcinoma, string carcinoma, sweat gland carcinoma, carcinoma telangiectaticum, carcinoma telangiectodes, transitional cell carcinoma, carcinoma tuberosum, tubular carcinoma, tuberous carcinoma, undifferentiated carcinoma, verrucous carcinoma, or carcinoma villosum.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat pancreatic cancer, breast cancer, multiple myeloma, cancers of secretory cells. For example certain methods herein treat cancer by decreasing or reducing or preventing the occurrence, growth, metastasis, or progression of cancer. In some embodiments, the methods described herein may be used to treat cancer by decreasing or eliminating a symptom of cancer. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a cancer described herein (e.g., pancreatic cancer, breast cancer, multiple myeloma, cancers of secretory cells).

In some embodiments, the compounds (compounds described herein, e.g., a compound of Formula (I)) and compositions (e.g., compositions comprising a compound described herein, e.g., a compound of Formula (I))) are used with a cancer immunotherapy (e.g., a checkpoint blocking antibody) to treat a subject (e.g., a human subject), e.g., suffering from a disease or disorder described herein (e.g., abnormal cell growth, e.g., cancer (e.g., a cancer described herein)). The methods described herein comprise administering a compound described herein, e.g., a compound of Formula (I) and an immunotherapy to a subject having abnormal cell growth such as cancer. Exemplary immunotherapies include, but are not limited to the following.

In some embodiments, the immunotherapeutic agent is a compound (e.g., a ligand, an antibody) that inhibits the immune checkpoint blockade pathway. In some embodiments, the immunotherapeutic agent is a compound that inhibits the indoleamine 2,3-dioxygenase (IDO) pathway. In some embodiments, the immunotherapeutic agent is a compound that agonizes the STING pathway. Cancer immunotherapy refers to the use of the immune system to treat cancer. Three groups of immunotherapy used to treat cancer include cell-based, antibody-based, and cytokine therapies. All groups exploit cancer cells' display of subtly different structures (e.g., molecular structure; antigens, proteins, molecules, carbohydrates) on their surface that can be detected by the immune system. Cancer immunotherapy (i.e., anti-tumor immunotherapy or anti-tumor immunotherapeutics) includes but is not limited to, immune checkpoint antibodies (e.g., PD-1 antibodies, PD-L1 antibodies, PD-L2 antibodies, CTLA-4 antibodies, TIM3 antibodies, LAG3 antibodies, TIGIT antibodies); and cancer vaccines (i.e., anti-tumor vaccines or vaccines based on neoantigens such as a peptide or RNA vaccine).

Cell-based therapies (e.g., cancer vaccines), usually involve the removal of immune cells from a subject suffering from cancer, either from the blood or from a tumor. Immune cells specific for the tumor will be activated, grown, and returned to a subject suffering from cancer where the immune cells provide an immune response against the cancer. Cell types that can be used in this way are e.g., natural killer cells, lymphokine-activated killer cells, cytotoxic T-cells, dendritic cells, CAR-T therapies (i.e., chimeric antigen receptor T-cells which are T-cells engineered to target specific antigens), TIL therapy (i.e., administration of tumor-infiltrating lymphocytes), TCR gene therapy, protein vaccines, and nucleic acid vaccines. An exemplary cell-based therapy is Provenge. In some embodiments, the cell-based therapy is a CAR-T therapy.

Interleukin-2 and interferon-alpha are examples of cytokines, proteins that regulate and coordinate the behavior of the immune system.

Cancer Vaccines with Neoantigens

Neoantigens are antigens encoded by tumor-specific mutated genes. Technological innovations have made it possible to dissect the immune response to patient-specific neoantigens that arise as a consequence of tumor-specific mutations, and emerging data suggest that recognition of such neoantigens is a major factor in the activity of clinical immunotherapies. These observations indicate that neoantigen load may form a biomarker in cancer immunotherapy. Many novel therapeutic approaches are being developed that selectively enhance T cell reactivity against this class of antigens. One approach to target neoantigens is via cancer vaccine. These vaccines can be developed using peptides or RNA, e.g., synthetic peptides or synthetic RNA.

Antibody therapies are antibody proteins produced by the immune system and that bind to a target antigen on the surface of a cell. Antibodies are typically encoded by an immunoglobulin gene or genes, or fragments thereof. In normal physiology antibodies are used by the immune system to fight pathogens. Each antibody is specific to one or a few proteins, and those that bind to cancer antigens are used, e.g., for the treatment of cancer. Antibodies are capable of specifically binding an antigen or epitope. (Fundamental Immunology, 3^(rd) Edition, W. E., Paul, ed., Raven Press, N.Y. (1993). Specific binding occurs to the corresponding antigen or epitope even in the presence of a heterogeneous population of proteins and other biologics. Specific binding of an antibody indicates that it binds to its target antigen or epitope with an affinity that is substantially greater than binding to irrelevant antigens. The relative difference in affinity is often at least 25% greater, more often at least 50% greater, most often at least 100% greater. The relative difference can be at least 2-fold, at least 5-fold, at least 10-fold, at least 25-fold, at least 50-fold, at least 100-fold, or at least 1000-fold, for example.

Exemplary types of antibodies include without limitation human, humanized, chimeric, monoclonal, polyclonal, single chain, antibody binding fragments, and diabodies. Once bound to a cancer antigen, antibodies can induce antibody-dependent cell-mediated cytotoxicity, activate the complement system, prevent a receptor interacting with its ligand or deliver a payload of chemotherapy or radiation, all of which can lead to cell death. Exemplary antibodies for the treatment of cancer include but are not limited to, Alemtuzumab, Bevacizumab, Bretuximab vedotin, Cetuximab, Gemtuzumab ozogamicin, Ibritumomab tiuxetan, Ipilimumab, Ofatumumab, Panitumumab, Rituximab, Tositumomab, Trastuzumab, Nivolumab, Pembrolizumab, Avelumab, durvalumab and pidilizumab.

Checkpoint Blocking Antibodies

The methods described herein comprise, in some embodiments, treating a human subject suffering from a disease or disorder described herein, the method comprising administering a composition comprising a cancer immunotherapy (e.g., an immunotherapeutic agent). In some embodiments, the immunotherapeutic agent is a compound (e.g., an inhibitor or antibody) that inhibits the immune checkpoint blockade pathway. Immune checkpoint proteins, under normal physiological conditions, maintain self-tolerance (e.g., prevent autoimmunity) and protect tissues from damage when the immune system is responding to e.g., pathogenic infection. Immune checkpoint proteins can be dysregulated by tumors as an important immune resistance mechanism. (Pardoll, Nature Rev. Cancer, 2012, 12, 252-264). Agonists of co-stimulatory receptors or antagonists of inhibitory signals (e.g., immune checkpoint proteins), provide an amplification of antigen-specific T-cell responses. Antibodies that block immune checkpoints do not target tumor cells directly but typically target lymphocyte receptors or their ligands to enhance endogenous antitumor activity.

Exemplary checkpoint blocking antibodies include but are not limited to, anti-CTLA-4, anti-PD-1, anti-LAG3 (i.e., antibodies against lymphocyte activation gene 3), and anti-TIM3 (i.e., antibodies against T-cell membrane protein 3). Exemplary anti-CTLA-4 antibodies include but are not limited to, ipilimumab and tremelimumab. Exemplary anti-PD-1 ligands include but are not limited to, PD-L1 (i.e., B7-H1 and CD274) and PD-L2 (i.e., B7-DC and CD273). Exemplary anti-PD-1 antibodies include but are not limited to, nivolumab (i.e., MDX-1106, BMS-936558, or ONO-4538)), CT-011, AMP-224, pembrolizumab (trade name Keytruda), and MK-3475. Exemplary PD-L1-specific antibodies include but are not limited to, BMS936559 (i.e., MDX-1105), MEDI4736 and MPDL-3280A. Exemplary checkpoint blocking antibodies also include but are not limited to, IMP321 and MGA271.

T-regulatory cells (e.g., CD4+, CD25+, or T-reg) are also involved in policing the distinction between self and non-self (e.g., foreign) antigens, and may represent an important mechanism in suppression of immune response in many cancers. T-reg cells can either emerge from the thymus “natural T-rest”) or can differentiate from mature T-cells under circumstances of peripheral tolerance induction “induced T-reg”). Strategies that minimize the action of T-reg cells would therefore be expected to facilitate the immune response to tumors. (Sutmuller, van Duivernvoorde et al., 2001).

IDO Pathway Inhibitors

The IDO pathway regulates immune response by suppressing T cell function and enabling local tumor immune escape. IDO expression by antigen-presenting cells (APCs) can lead to tryptophan depletion, and resulting antigen-specific T cell energy and regulatory T cell recruitment. Some tumors even express IDO to shield themselves from the immune system. A compound that inhibits IDO or the IDO pathway thereby activating the immune system to attack the cancer (e.g., tumor in a subject). Exemplary IDO pathway inhibitors include indoximod, epacadostat and EOS200271.

STING Pathway Agonists

Stimulator of interferon genes (STING) is an adaptor protein that plays an important role in the activation of type I interferons in response to cytosolic nucleic acid ligands. Evidence indicates involvement of the STING pathway in the induction of antitumor immune response. It has been shown that activation of the STING-dependent pathway in cancer cells can result in tumor infiltration with immune cells and modulation of the anticancer immune response. STING agonists are being developed as a class of cancer therapeutics. Exemplary STING agonists include MK-1454 and ADU-S100.

Co-Stimulatory Antibodies

The methods described herein comprise, in some embodiments, treating a human subject suffering from a disease or disorder described herein, the method comprising administering a composition comprising a cancer immunotherapy (e.g., an immunotherapeutic agent). In some embodiments, the immunotherapeutic agent is a co-stimulatory inhibitor or antibody. In some embodiments, the methods described herein comprise depleting or activating anti-4-1BB, anti-OX40, anti-GITR, anti-CD27 and anti-CD40, and variants thereof.

Inventive methods of the present invention contemplate single as well as multiple administrations of a therapeutically effective amount of a compound as described herein. Compounds, e.g., a compound as described herein, can be administered at regular intervals, depending on the nature, severity and extent of the subject's condition. In some embodiments, a compound described herein is administered in a single dose. In some embodiments, a compound described herein is administered in multiple doses.

Inflammatory Disease

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat an inflammatory disease. As used herein, the term “inflammatory disease” refers to a disease or condition characterized by aberrant inflammation (e.g. an increased level of inflammation compared to a control such as a healthy person not suffering from a disease). Examples of inflammatory diseases include postoperative cognitive dysfunction, arthritis (e.g., rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis), systemic lupus erythematosus (SLE), myasthenia gravis, juvenile onset diabetes, diabetes mellitus type 1, Guillain-Barre syndrome, Hashimoto's encephalitis, Hashimoto's thyroiditis, ankylosing spondylitis, psoriasis, Sjogren's syndrome, vasculitis, glomerulonephritis, auto-immune thyroiditis, Behcet's disease, Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, ichthyosis, Graves' ophthalmopathy, inflammatory bowel disease, Addison's disease, Vitiligo, asthma (e.g., allergic asthma), acne vulgaris, celiac disease, chronic prostatitis, inflammatory bowel disease, pelvic inflammatory disease, reperfusion injury, sarcoidosis, transplant rejection, interstitial cystitis, atherosclerosis, and atopic dermatitis. Proteins associated with inflammation and inflammatory diseases (e.g. aberrant expression being a symptom or cause or marker of the disease) include interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-18 (IL-18), TNF-α (tumor necrosis factor-alpha), and C-reactive protein (CRP).

In some embodiments, the inflammatory disease comprises postoperative cognitive dysfunction, arthritis (e.g., rheumatoid arthritis, psoriatic arthritis, or juvenile idiopathic arthritis), systemic lupus erythematosus (SLE), myasthenia gravis, diabetes (e.g., juvenile onset diabetes or diabetes mellitus type 1), Guillain-Barre syndrome, Hashimoto's encephalitis, Hashimoto's thyroiditis, ankylosing spondylitis, psoriasis, Sjogren's syndrome, vasculitis, glomerulonephritis, auto-immune thyroiditis, Behcet's disease, Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, ichthyosis, Graves' ophthalmopathy, inflammatory bowel disease, Addison's disease, vitiligo, asthma (e.g., allergic asthma), acne vulgaris, celiac disease, chronic prostatitis, pelvic inflammatory disease, reperfusion injury, sarcoidosis, transplant rejection, interstitial cystitis, atherosclerosis, or atopic dermatitis.

In some embodiments, the inflammatory disease comprises postoperative cognitive dysfunction, which refers to a decline in cognitive function (e.g. memory or executive function (e.g. working memory, reasoning, task flexibility, speed of processing, or problem solving)) following surgery.

In other embodiments, the method of treatment is a method of prevention. For example, a method of treating postsurgical cognitive dysfunction may include preventing postsurgical cognitive dysfunction or a symptom of postsurgical cognitive dysfunction or reducing the severity of a symptom of postsurgical cognitive dysfunction by administering a compound described herein prior to surgery.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat an inflammatory disease (e.g., an inflammatory disease described herein) by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat an inflammatory disease (e.g., an inflammatory disease described herein).

Musculoskeletal Diseases

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a musculoskeletal disease. As used herein, the term “musculoskeletal disease” refers to a disease or condition in which the function of a subject's musculoskeletal system (e.g., muscles, ligaments, tendons, cartilage, or bones) becomes impaired. Exemplary musculoskeletal diseases that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include muscular dystrophy (e.g., Duchenne muscular dystrophy, Becker muscular dystrophy, distal muscular dystrophy, congenital muscular dystrophy, Emery-Dreifuss muscular dystrophy, facioscapulohumeral muscular dystrophy, myotonic muscular dystrophy type 1, or myotonic muscular dystrophy type 2), limb girdle muscular dystrophy, multisystem proteinopathy, rhizomelic chondrodysplasia punctata, X-linked recessive chondrodysplasia punctata, Conradi-Hünermann syndrome, Autosomal dominant chondrodysplasia punctata, stress induced skeletal disorders (e.g., stress induced osteoporosis), multiple sclerosis, amyotrophic lateral sclerosis (ALS), primary lateral sclerosis, progressive muscular atrophy, progressive bulbar palsy, pseudobulbar palsy, spinal muscular atrophy, progressive spinobulbar muscular atrophy, spinal cord spasticity, spinal muscle atrophy, myasthenia gravis, neuralgia, fibromyalgia, Machado-Joseph disease, Paget's disease of bone, cramp fasciculation syndrome, Freidrich's ataxia, a muscle wasting disorder (e.g., muscle atrophy, sarcopenia, cachexia), an inclusion body myopathy, motor neuron disease, or paralysis.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a musculoskeletal disease (e.g., a musculoskeletal disease described herein) by decreasing or eliminating a symptom of the disease. In some embodiments, the method of treatment comprises treatment of muscle pain or muscle stiffness associated with a musculoskeletal disease. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a musculoskeletal disease (e.g., a musculoskeletal disease described herein).

Metabolic Diseases

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat metabolic disease. As used herein, the term “metabolic disease” refers to a disease or condition affecting a metabolic process in a subject. Exemplary metabolic diseases that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), liver fibrosis, obesity, heart disease, atherosclerosis, arthritis, cystinosis, diabetes (e.g., Type I diabetes, Type II diabetes, or gestational diabetes), phenylketonuria, proliferative retinopathy, or Kearns-Sayre disease.

In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a metabolic disease (e.g., a metabolic disease described herein) by decreasing or eliminating a symptom of the disease. In some embodiments, the method of treatment comprises decreasing or eliminating a symptom comprising elevated blood pressure, elevated blood sugar level, weight gain, fatigue, blurred vision, abdominal pain, flatulence, constipation, diarrhea, jaundice, and the like. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a metabolic disease (e.g., a musculoskeletal disease described herein).

Mitochondrial Diseases

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat mitochondrial disease. As used herein, the term “mitochondrial disease” refers to a disease or condition affecting the mitochondria in a subject. In some embodiments, the mitochondrial disease is associated with, or is a result of, or is caused by mitochondrial dysfunction, one or more mitochondrial protein mutations, or one or more mitochondrial DNA mutations. In some embodiments, the mitochondrial disease is a mitochondrial myopathy. In some embodiments, mitochondrial diseases, e.g., the mitochondrial myopathy, that may be treated with a compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include, e.g., Barth syndrome, chronic progressive external ophthalmoplegia (cPEO), Kearns-Sayre syndrome (KSS), Leigh syndrome (e.g., MILS, or maternally inherited Leigh syndrome), mitochondrial DNA depletion syndromes (MDDS, e.g., Alpers syndrome), mitochondrial encephalomyopathy (e.g., mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS)), mitochondrial neurogastrointestinal encephalomyopathy (MNGIE), myoclonus epilepsy with ragged red fibers (MERRF), neuropathy, ataxia, retinitis pigmentosa (NARP), Leber's hereditary optic neuropathy (LHON), and Pearson syndrome.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a mitochondrial disease described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a mitochondrial disease described herein.

Hearing Loss

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat hearing loss. As used herein, the term “hearing loss” or “hearing loss condition” may broadly encompass any damage to the auditory systems, organs, and cells or any impairment of an animal subject's ability to hear sound, as measured by standard methods and assessments known in the art, for example otoacoustic emission testing, pure tone testing, and auditory brainstem response testing. Exemplary hearing loss conditions that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include, but are not limited to, mitochondrial nonsyndromic hearing loss and deafness, hair cell death, age-related hearing loss, noise-induced hearing loss, genetic or inherited hearing loss, hearing loss experienced as a result of ototoxic exposure, hearing loss resulting from disease, and hearing loss resulting from trauma. In some embodiments, mitochondrial nonsyndromic hearing loss and deafness is a MT-RNR1-related hearing loss. In some embodiments, the MT-RNR1-related hearing loss is the result of amino glycoside ototoxicity. In some embodiments, mitochondrial nonsyndromic hearing loss and deafness is a MT-TS1-related hearing loss. In some embodiments, mitochondrial nonsyndromic hearing loss and deafness is characterized by sensorineural hearing loss.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a hearing loss condition described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a hearing loss condition described herein.

Ocular Disease

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat eye disease. As used herein, the term “ocular disease” may refer to a disease or condition in which the function of a subject's eye becomes impaired. Exemplary ocular diseases and conditions that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include cataracts, glaucoma, endoplasmic reticulum (ER) stress, autophagy deficiency, age-related macular degeneration (AMD), or diabetic retinopathy.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat an ocular disease or condition described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat an ocular disease or condition described herein.

Kidney Diseases

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat kidney disease. As used herein, the term “kidney disease” may refer to a disease or condition in which the function of a subject's kidneys becomes impaired. Exemplary kidney diseases that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include Abderhalden KaufmannLignac syndrome (Nephropathic Cystinosis), Abdominal Compartment Syndrome, Acetaminophen-induced Nephrotoxicity, Acute Kidney Failure/Acute Kidney Injury, Acute Lobar Nephronia, Acute Phosphate Nephropathy, Acute Tubular Necrosis, Adenine Phosphoribosyltransferase Deficiency, Adenovirus Nephritis, Alagille Syndrome, Alport Syndrome, Amyloidosis, ANCA Vasculitis Related to Endocarditis and Other Infections, Angiomyolipoma, Analgesic Nephropathy, Anorexia Nervosa and Kidney Disease, Angiotensin Antibodies and Focal Segmental Glomerulosclerosis, Antiphospholipid Syndrome, Anti-TNF-α Therapy-related Glomerulonephritis, APOL1 Mutations, Apparent Mineralocorticoid Excess Syndrome, Aristolochic Acid Nephropathy, Chinese Herbal Nephropathy, Balkan Endemic Nephropathy, Arteriovenous Malformations and Fistulas of the Urologic Tract, Autosomal Dominant Hypocalcemia, Bardet-Biedl Syndrome, Bartter Syndrome, Bath Salts and Acute Kidney Injury, Beer Potomania, Beeturia, β-Thalassemia Renal Disease, Bile Cast Nephropathy, BK Polyoma Virus Nephropathy in the Native Kidney, Bladder Rupture, Bladder Sphincter Dyssynergia, Bladder Tamponade, Border-Crossers' Nephropathy, Bourbon Virus and Acute Kidney Injury, Burnt Sugarcane Harvesting and Acute Renal Dysfunction, Byetta and Renal Failure, C1q Nephropathy, C3 Glomerulopathy, C3 Glomerulopathy with Monoclonal Gammopathy, C4 Glomerulopathy, Calcineurin Inhibitor Nephrotoxicity, Callilepsis Laureola Poisoning, Cannabinoid Hyperemesis Acute Renal Failure, Cardiorenal syndrome, Carfilzomib-Indiced Renal Injury, CFHR5 nephropathy, CharcotMarieTooth Disease with Glomerulopathy, Chinese Herbal Medicines and Nephrotoxicity, Cherry Concentrate and Acute Kidney Injury, Cholesterol Emboli, ChurgStrauss syndrome, Chyluria, Ciliopathy, Cocaine and the Kidney, Cold Diuresis, Colistin Nephrotoxicity, Collagenofibrotic Glomerulopathy, Collapsing Glomerulopathy, Collapsing Glomerulopathy Related to CMV, Combination Antiretroviral (cART) Related-Nephropathy, Congenital Anomalies of the Kidney and Urinary Tract (CAKUT), Congenital Nephrotic Syndrome, Congestive Renal Failure, Conorenal syndrome (Mainzer-Saldino Syndrome or Saldino-Mainzer Disease), Contrast Nephropathy, Copper Sulphate Intoxication, Cortical Necrosis, Crizotinib-related Acute Kidney Injury, Cryocrystalglobulinemia, Cryoglobuinemia, Crystalglobulin-Induced Nephropathy, Crystal-Induced Acute Kidney injury, Crystal-Storing Histiocytosis, Cystic Kidney Disease, Acquired, Cystinuria, Dasatinib-Induced Nephrotic-Range Proteinuria, Dense Deposit Disease (MPGN Type 2), Dent Disease (X-linked Recessive Nephrolithiasis), DHA Crystalline Nephropathy, Dialysis Disequilibrium Syndrome, Diabetes and Diabetic Kidney Disease, Diabetes Insipidus, Dietary Supplements and Renal Failure, Diffuse Mesangial Sclerosis, Diuresis, Djenkol Bean Poisoning (Djenkolism), Down Syndrome and Kidney Disease, Drugs of Abuse and Kidney Disease, Duplicated Ureter, EAST syndrome, Ebola and the Kidney, Ectopic Kidney, Ectopic Ureter, Edema, Swelling, Erdheim-Chester Disease, Fabry's Disease, Familial Hypocalciuric Hypercalcemia, Fanconi Syndrome, Fraser syndrome, Fibronectin Glomerulopathy, Fibrillary Glomerulonephritis and Immunotactoid Glomerulopathy, Fraley syndrome, Fluid Overload, Hypervolemia, Focal Segmental Glomerulosclerosis, Focal Sclerosis, Focal Glomerulosclerosis, Galloway Mowat syndrome, Giant Cell (Temporal) Arteritis with Kidney Involvement, Gestational Hypertension, Gitelman Syndrome, Glomerular Diseases, Glomerular Tubular Reflux, Glycosuria, Goodpasture Syndrome, Green Smoothie Cleanse Nephropathy, HANAC Syndrome, Harvoni (Ledipasvir with Sofosbuvir)-Induced Renal Injury, Hair Dye Ingestion and Acute Kidney Injury, Hantavirus Infection Podocytopathy, Heat Stress Nephropathy, Hematuria (Blood in Urine), Hemolytic Uremic Syndrome (HUS), Atypical Hemolytic Uremic Syndrome (aHUS), Hemophagocytic Syndrome, Hemorrhagic Cystitis, Hemorrhagic Fever with Renal Syndrome (HFRS, Hantavirus Renal Disease, Korean Hemorrhagic Fever, Epidemic Hemorrhagic Fever, Nephropathis Epidemica), Hemosiderinuria, Hemosiderosis related to Paroxysmal Nocturnal Hemoglobinuria and Hemolytic Anemia, Hepatic Glomerulopathy, Hepatic Veno-Occlusive Disease, Sinusoidal Obstruction Syndrome, Hepatitis C-Associated Renal Disease, Hepatocyte Nuclear Factor 1β-Associated Kidney Disease, Hepatorenal Syndrome, Herbal Supplements and Kidney Disease, High Altitude Renal Syndrome, High Blood Pressure and Kidney Disease, HIV-Associated Immune Complex Kidney Disease (HIVICK), HIV-Associated Nephropathy (HIVAN), HNF1B-related Autosomal Dominant Tubulointerstitial Kidney Disease, Horseshoe Kidney (Renal Fusion), Hunner's Ulcer, Hydroxychloroquine-induced Renal Phospholipidosis, Hyperaldosteronism, Hypercalcemia, Hyperkalemia, Hypermagnesemia, Hypernatremia, Hyperoxaluria, Hyperphosphatemia, Hypocalcemia, Hypocomplementemic Urticarial Vasculitic Syndrome, Hypokalemia, Hypokalemia-induced renal dysfunction, Hypokalemic Periodic Paralysis, Hypomagnesemia, Hyponatremia, Hypophosphatemia, Hypophosphatemia in Users of Cannabis, Hypertension, Hypertension, Monogenic, Iced Tea Nephropathy, Ifosfamide Nephrotoxicity, IgA Nephropathy, IgG4 Nephropathy, Immersion Diuresis, Immune-Checkpoint Therapy-Related Interstitial Nephritis, Infliximab-Related Renal Disease, Interstitial Cystitis, Painful Bladder Syndrome (Questionnaire), Interstitial Nephritis, Interstitial Nephritis, Karyomegalic, Ivemark's syndrome, JC Virus Nephropathy, Joubert Syndrome, Ketamine-Associated Bladder Dysfunction, Kidney Stones, Nephrolithiasis, Kombucha Tea Toxicity, Lead Nephropathy and Lead-Related Nephrotoxicity, Lecithin Cholesterol Acyltransferase Deficiency (LCAT Deficiency), Leptospirosis Renal Disease, Light Chain Deposition Disease, Monoclonal Immunoglobulin Deposition Disease, Light Chain Proximal Tubulopathy, Liddle Syndrome, Lightwood-Albright Syndrome, Lipoprotein Glomerulopathy, Lithium Nephrotoxicity, LMX1B Mutations Cause Hereditary FSGS, Loin Pain Hematuria, Lupus, Systemic Lupus Erythematosis, Lupus Kidney Disease, Lupus Nephritis, Lupus Nephritis with Antineutrophil Cytoplasmic Antibody Seropositivity, Lupus Podocytopathy, Lyme Disease-Associated Glomerulonephritis, Lysinuric Protein Intolerance, Lysozyme Nephropathy, Malarial Nephropathy, Malignancy-Associated Renal Disease, Malignant Hypertension, Malakoplakia, McKittrick-Wheelock Syndrome, MDMA (Molly; Ecstacy; 3,4-Methylenedioxymethamphetamine) and Kidney Failure, Meatal Stenosis, Medullary Cystic Kidney Disease, Urolodulin-Associated Nephropathy, Juvenile Hyperuricemic Nephropathy Type 1, Medullary Sponge Kidney, Megaureter, Melamine Toxicity and the Kidney, MELAS Syndrome, Membranoproliferative Glomerulonephritis, Membranous Nephropathy, Membranous-like Glomerulopathy with Masked IgG Kappa Deposits, MesoAmerican Nephropathy, Metabolic Acidosis, Metabolic Alkalosis, Methotrexate-related Renal Failure, Microscopic Polyangiitis, Milk-alkalai syndrome, Minimal Change Disease, Monoclonal Gammopathy of Renal Significance, Dysproteinemia, Mouthwash Toxicity, MUC1 Nephropathy, Multicystic dysplastic kidney, Multiple Myeloma, Myeloproliferative Neoplasms and Glomerulopathy, Nail-patella Syndrome, NARP Syndrome, Nephrocalcinosis, Nephrogenic Systemic Fibrosis, Nephroptosis (Floating Kidney, Renal Ptosis), Nephrotic Syndrome, Neurogenic Bladder, 9/11 and Kidney Disease, Nodular Glomerulosclerosis, Non-Gonococcal Urethritis, Nutcracker syndrome, Oligomeganephronia, Orofaciodigital Syndrome, Orotic Aciduria, Orthostatic Hypotension, Orthostatic Proteinuria, Osmotic Diuresis, Osmotic Nephrosis, Ovarian Hyperstimulation Syndrome, Oxalate Nephropathy, Page Kidney, Papillary Necrosis, Papillorenal Syndrome (Renal-Coloboma Syndrome, Isolated Renal Hypoplasia), PARN Mutations and Kidney Disease, Parvovirus B19 and the Kidney, The Peritoneal-Renal Syndrome, POEMS Syndrome, Posterior Urethral Valve, Podocyte Infolding Glomerulopathy, Post-infectious Glomerulonephritis, Post-streptococcal Glomerulonephritis, Post-infectious Glomerulonephritis, Atypical, Post-Infectious Glomerulonephritis (IgA-Dominant), Mimicking IgA Nephropathy, Polyarteritis Nodosa, Polycystic Kidney Disease, Posterior Urethral Valves, Post-Obstructive Diuresis, Preeclampsia, Propofol infusion syndrome, Proliferative Glomerulonephritis with Monoclonal IgG Deposits (Nasr Disease), Propolis (Honeybee Resin) Related Renal Failure, Proteinuria (Protein in Urine), Pseudohyperaldosteronism, Pseudohypobicarbonatemia, Pseudohypoparathyroidism, Pulmonary-Renal Syndrome, Pyelonephritis (Kidney Infection), Pyonephrosis, Pyridium and Kidney Failure, Radiation Nephropathy, Ranolazine and the Kidney, Refeeding syndrome, Reflux Nephropathy, Rapidly Progressive Glomerulonephritis, Renal Abscess, Peripnephric Abscess, Renal Agenesis, Renal Arcuate Vein Microthrombi-Associated Acute Kidney Injury, Renal Artery Aneurysm, Renal Artery Dissection, Spontaneous, Renal Artery Stenosis, Renal Cell Cancer, Renal Cyst, Renal Hypouricemia with Exercise-induced Acute Renal Failure, Renal Infarction, Renal Osteodystrophy, Renal Tubular Acidosis, Renin Mutations and Autosomal Dominant Tubulointerstitial Kidney Disease, Renin Secreting Tumors (Juxtaglomerular Cell Tumor), Reset Osmostat, Retrocaval Ureter, Retroperitoneal Fibrosis, Rhabdomyolysis, Rhabdomyolysis related to Bariatric Sugery, Rheumatoid Arthritis-Associated Renal Disease, Sarcoidosis Renal Disease, Salt Wasting, Renal and Cerebral, Schistosomiasis and Glomerular Disease, Schimke immuno-osseous dysplasia, Scleroderma Renal Crisis, Serpentine Fibula-Polycystic Kidney Syndrome, Exner Syndrome, Sickle Cell Nephropathy, Silica Exposure and Chronic Kidney Disease, Sri Lankan Farmers' Kidney Disease, Sjögren's Syndrome and Renal Disease, Synthetic Cannabinoid Use and Acute Kidney Injury, Kidney Disease Following Hematopoietic Cell Transplantation, Kidney Disease Related to Stem Cell Transplantation, TAFRO Syndrome, Tea and Toast Hyponatremia, Tenofovir-Induced Nephrotoxicity, Thin Basement Membrane Disease, Benign Familial Hematuria, Thrombotic Microangiopathy Associated with Monoclonal Gammopathy, Trench Nephritis, Trigonitis, Tuberculosis, Genitourinary, Tuberous Sclerosis, Tubular Dysgenesis, Immune Complex Tubulointerstitial Nephritis Due to Autoantibodies to the Proximal Tubule Brush Border, Tumor Lysis Syndrome, Uremia, Uremic Optic Neuropathy, Ureteritis Cystica, Ureterocele, Urethral Caruncle, Urethral Stricture, Urinary Incontinence, Urinary Tract Infection, Urinary Tract Obstruction, Urogenital Fistula, Uromodulin-Associated Kidney Disease, Vancomycin-Associated Cast Nephropathy, Vasomotor Nephropathy, Vesicointestinal Fistula, Vesicoureteral Reflux, VGEF Inhibition and Renal Thrombotic Microangiopathy, Volatile Anesthetics and Acute Kidney Injury, Von Hippel-Lindau Disease, Waldenstrom's Macroglobulinemic Glomerulonephritis, Warfarin-Related Nephropathy, Wasp Stings and Acute Kidney Injury, Wegener's Granulomatosis, Granulomatosis with Polyangiitis, West Nile Virus and Chronic Kidney Disease, Wunderlich syndrome, Zellweger Syndrome, or Cerebrohepatorenal Syndrome.

Infectious Diseases

In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I) is used to treat an infectious disease. Exemplary infectious diseases that may be treated with a compound disclosed herein, e.g., a compound of Formula include bacterial infections, viral infections (e.g., herpes, shingles, influenza, the common cold, encephalitis), and parasitic infections.

In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I) is used to treat an infectious disease (e.g., an infectious disease described herein) by decreasing or eliminating a symptom of the disease. In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I) may be used as a single agent in a composition or in combination with another agent in a composition to treat an infectious disease.

Parasitic Infections

In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I) is used to treat a parasitic infection.

In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I) is used to treat a parasitic infection by decreasing or eliminating a symptom of the disease. In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I) may be used as a single agent in a composition or in combination with another agent in a composition to treat a parasitic infection.

Immunosuppressive Diseases

In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I) is used to treat an immunosuppressive disease.

In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I) is used to treat an immunosuppressive disease by decreasing or eliminating a symptom of the disease. In some embodiments, a compound disclosed herein, e.g., a compound of Formula (I) may be used as a single agent in a composition or in combination with another agent in a composition to treat a immunosuppressive disease.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a kidney disease described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a kidney disease described herein.

Skin Diseases

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a skin disease. As used herein, the term “skin disease” may refer to a disease or condition affecting the skin. Exemplary skin diseases that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include acne, alopecia areata, basal cell carcinoma, Bowen's disease, congenital erythropoietic porphyria, contact dermatitis, Darier's disease, disseminated superficial actinic porokeratosis, dystrophic epidermolysis bullosa, eczema (atopic eczema), extra-mammary Paget's disease, epidermolysis bullosa simplex, erythropoietic protoporphyria, fungal infections of nails, Hailey-Hailey disease, herpes simplex, hidradenitis suppurativa, hirsutism, hyperhidrosis, ichthyosis, impetigo, keloids, keratosis pilaris, lichen planus, lichen sclerosus, melanoma, melasma, mucous membrane pemphigoid, pemphigoid, pemphigus vulgaris, pityriasis lichenoides, pityriasis rubra pilaris, plantar warts (verrucas), polymorphic light eruption, psoriasis, plaque psoriasis, pyoderma gangrenosum, rosacea, scabies, scleroderma, shingles, squamous cell carcinoma, sweet's syndrome, urticaria and angioedema and vitiligo.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a skin disease described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a skin disease described herein.

Fibrotic Diseases

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a fibrotic disease. As used herein, the term “fibrotic disease” may refer to a disease or condition that is defined by the accumulation of excess extracellular matrix components. Exemplary fibrotic diseases that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include adhesive capsulitis, arterial stiffness, arthrofibrosis, atrial fibrosis, cardiac fibrosis, cirrhosis, congenital hepatic fibrosis, Crohn's disease, cystic fibrosis, Dupuytren's contracture, endomyocardial fibrosis, glial scar, hepatitis C, hypertrophic cardiomyopathy, hypersensitivity pneumonitis, idiopathic pulmonary fibrosis, idiopathic interstitial pneumonia, interstitial lung disease, keloid, mediastinal fibrosis, myelofibrosis, nephrogenic systemic fibrosis, non-alcoholic fatty liver disease, old myocardial infarction, Peyronie's disease, pneumoconiosis, pneumonitis, progressive massive fibrosis, pulmonary fibrosis, radiation-induced lung injury, retroperitoneal fibrosis, scleroderma/systemic sclerosis, silicosis and ventricular remodeling.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a fibrotic disease described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a fibrotic disease described herein.

Hemoglobin Disorders

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a hemoglobin disease. As used herein, the terms “hemoglobin disease” or “hemoglobin disorder” may refer to a disease or condition characterized by an abnormal production or structure of the hemoglobin protein. Exemplary hemoglobin diseases that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include “dominant” β-thalassemia, acquired (toxic) methemoglobinemia, carboxyhemoglobinemia, congenital Heinz body hemolytic anemia, HbH disease, HbS/β-thalassemia, HbE/β-thalassemia, HbSC disease, homozygous α⁺-thalassemia (phenotype of α⁰-thalassemia), Hydrops fetalis with Hb Bart's, sickle cell anemia/disease, sickle cell trait, sickle β-thalassemia disease, α⁺-thalassemia, α⁰-thalassemia, α-Thalassemia associated with myelodysplastic syndromes, α-Thalassemia with mental retardation syndrome (ATR), β⁰-Thalassemia, β⁺-Thalassemia, δ-Thalassemia, γ-Thalassemia, β-Thalassemia major, β-Thalassemia intermedia, δβ-Thalassemia, and εγδβ-Thalassemia.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a hemoglobin disease described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a hemoglobin disease described herein.

Autoimmune Diseases

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat an autoimmune disease. As used herein, the term “autoimmune disease” may refer to a disease or condition in which the immune system of a subject attacks and damages the tissues of said subject. Exemplary kidney diseases that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include Achalasia, Addison's disease, Adult Still's disease, Agammaglobulinemia, Alopecia areata, Amyloidosis, Ankylosing spondylitis, Anti-GBM/Anti-TBM nephritis, Antiphospholipid syndrome, Autoimmune angioedema, Autoimmune dysautonomia, Autoimmune encephalomyelitis, Autoimmune hepatitis, Autoimmune inner ear disease (AIED), Autoimmune myocarditis, Autoimmune oophoritis, Autoimmune orchitis, Autoimmune pancreatitis, Autoimmune retinopathy, Autoimmune urticaria, Axonal & neuronal neuropathy (AMAN), Bal disease, Behcet's disease, Benign mucosal pemphigoid, Bullous pemphigoid, Castleman disease (CD), Celiac disease, Chagas disease, Chronic inflammatory demyelinating polyneuropathy (CIDP), Chronic recurrent multifocal osteomyelitis (CRMO), Churg-Strauss Syndrome (CSS) or Eosinophilic Granulomatosis (EGPA), Cicatricial pemphigoid, Cogan's syndrome, Cold agglutinin disease, Congenital heart block, Coxsackie myocarditis, CREST syndrome, Crohn's disease, Dermatitis herpetiformis, Dermatomyositis, Devic's disease (neuromyelitis optica), Discoid lupus, Dressler's syndrome, Endometriosis, Eosinophilic esophagitis (EoE), Eosinophilic fasciitis, Erythema nodosum, Essential mixed cryoglobulinemia, Evans syndrome, Fibromyalgia, Fibrosing alveolitis, Giant cell arteritis (temporal arteritis), Giant cell myocarditis, Glomerulonephritis, Goodpasture's syndrome, Granulomatosis with Polyangiitis, Graves' disease, Guillain-Barre syndrome, Hashimoto's thyroiditis, Hemolytic anemia, Henoch-Schonlein purpura (HSP), Herpes gestationis or pemphigoid gestationis (PG), Hidradenitis Suppurativa (HS) (Acne Inversa), Hypogammalglobulinemia, IgA Nephropathy, IgG4-related sclerosing disease, Immune thrombocytopenic purpura (ITP), Inclusion body myositis (IBM), Interstitial cystitis (IC), Juvenile arthritis, Juvenile diabetes (Type 1 diabetes), Juvenile myositis (JM), Kawasaki disease, Lambert-Eaton syndrome, Leukocytoclastic vasculitis, Lichen planus, Lichen sclerosus, Ligneous conjunctivitis, Linear IgA disease (LAD), Lupus, Lyme disease chronic, Meniere's disease, Microscopic polyangiitis (MPA), Mixed connective tissue disease (MCTD), Mooren's ulcer, Mucha-Habermann disease, Multifocal Motor Neuropathy (MMN) or MMNCB, Multiple sclerosis, Myasthenia gravis, Myositis, Narcolepsy, Neonatal Lupus, Neuromyelitis optica, Neutropenia, Ocular cicatricial pemphigoid, Optic neuritis, Palindromic rheumatism (PR), PANDAS, Paraneoplastic cerebellar degeneration (PCD), Paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg syndrome, Pars planitis (peripheral uveitis), Parsonnage-Turner syndrome Pemphigus, Peripheral neuropathy, Perivenous encephalomyelitis, Pernicious anemia (PA), POEMS syndrome, Polyarteritis nodosa, Polyglandular syndrome type I, Polyglandular syndrome type II, Polyglandular syndrome type III, Polymyalgia rheumatica, Polymyositis, Postmyocardial infarction syndrome, Postpericardiotomy syndrome, Primary biliary cirrhosis, Primary sclerosing cholangitis, Progesterone dermatitis, Psoriasis, Psoriatic arthritis, Pure red cell aplasia (PRCA), Pyoderma gangrenosum, Raynaud's phenomenon, Reactive Arthritis, Reflex sympathetic dystrophy, Relapsing polychondritis, Restless legs syndrome (RLS), Retroperitoneal fibrosis, Rheumatic fever, Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome, Scleritis, Scleroderma, Sjögren's syndrome, Sperm & testicular autoimmunity, Stiff person syndrome (SPS), Subacute bacterial endocarditis (SBE), Susac's syndrome, Sympathetic ophthalmia (SO), Takayasu's arteritis, Temporal arteritis/Giant cell arteritis, Thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome (THS), Transverse myelitis, Type 1 diabetes, Ulcerative colitis (UC), Undifferentiated connective tissue disease (UCTD), Uveitis, Vasculitis, Vitiligo, Vogt-Koyanagi-Harada Disease, and Wegener's granulomatosis (or Granulomatosis with Polyangiitis (GPA)).

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat an autoimmune disease described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat an autoimmune disease described herein.

Viral Infections

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a viral infection. Exemplary viral infections that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include influenza, human immunodeficiency virus (HIV) and herpes.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a viral infection described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a viral infection described herein.

Malaria Infection

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a malaria. As used herein, the term “malaria” may refer to a parasitic disease of protozoan of the plasmodium genus that causes infection of red blood cells (RBCs). Exemplary forms of malaria infection that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof include infection caused by Plasmodium vivax, Plasmodium ovale, Plasmodium malariae and Plasmodium falciparum. In some embodiments, the malaria infection that may be treated with a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is resistant/recrudescent malaria.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a malaria infection described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a malaria infection described herein.

Diseases with Mutations Leading to Unfolded Protein Response (UPR) Induction

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a disease with mutations that leads to UPR induction. Exemplary disease with mutations that lead to UPR induction include Marinesco-Sjogren syndrome, neuropathic pain, diabetic neuropathic pain, noise induced hearing loss, non-syndromic sensorineural hearing loss, age-related hearing loss, Wolfram syndrome, Darier White disease, Usher syndrome, collagenopathies, Thin basement nephropathy, Alport syndrome, skeletal chondrodysplasia, metaphyseal chondrodysplasia type Schmid, and Pseudochondrodysplasia.

In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof is used to treat a disease with mutations that leads to UPR induction described herein by decreasing or eliminating a symptom of the disease. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be used as a single agent in a composition or in combination with another agent in a composition to treat a disease with mutations that leads to UPR induction described herein.

Methods of Modulating Protein Production

In another aspect, disclosed herein is a method of modulating the expression of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in a cell, the method comprising contacting the cell with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, thereby modulating the expression of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the cell. In some embodiments, contacting the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof with the cell increases the expression of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the cell. In some embodiments, contacting the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof with the cell decreases the expression of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the cell.

In another aspect, disclosed herein is a method of preventing or treating a condition, disease or disorder described herein in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, wherein the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof modulates the expression of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof by the patient's cells, thereby treating the condition, disease or disorder. In some embodiments, the condition, disease or disorder is characterized by aberrant expression of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof by the patient's cells. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases the expression of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof by the patient's cells, thereby treating the condition, disease or disorder. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof decreases the expression of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof by the patient's cells, thereby treating the condition, disease or disorder.

In another aspect, disclosed herein is a method of modulating the activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in a cell, the method comprising contacting the cell with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, thereby modulating the activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the cell. In some embodiments, contacting the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof with the cell increases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the cell. In some embodiments, contacting the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof with the cell decreases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the cell.

In another aspect, disclosed herein is a method of preventing or treating a condition, disease or disorder described herein in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, wherein the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof modulates the activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof by the patients cells, thereby treating the condition, disease or disorder. In some embodiments, the condition, disease or disorder is characterized by aberrant activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the patient's cells. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the patient's cells, thereby treating the condition, disease or disorder. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof decreases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the patient's cells, thereby treating the condition, disease or disorder.

In some embodiments, administering an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, wherein the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof modulates both the expression and the activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the patients cells, thereby treating the condition, disease or disorder.

In some embodiments, the compound of Formula (I) is chemically modified, prior to (ex vivo) or after (in vivo) contacting with a cell, forming a biologically active compound that modulates the expression and/or activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the cell. In some embodiments, the compound of Formula (I) is metabolized by the patient forming a biologically active compound that modulates the expression and/or activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the patients cells, thereby treating a condition, disease or disorder disclosed herein. In some embodiments, the biologically active compound is the compound of formula (II).

In one aspect, disclosed herein is a method of treating a disease related to a modulation of eIF2B activity or levels, eIF2α activity or levels, or the activity or levels of a component of the eIF2 pathway or the ISR pathway in a patient in need thereof, comprising administering to the patient an effective amount of a compound of Formula (I). In some embodiments, the modulation comprises an increase in eIF2B activity or levels, increase in eIF2α activity or levels, or increase in activity or levels of a component of the eIF2 pathway or the ISR pathway. In some embodiments, the disease may be caused by a mutation to a gene or protein sequence related to a member of the eIF2 pathway (e.g., the eIF2α signaling pathway).

Methods of Increasing Protein Activity and Production

In another aspect, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be useful in applications where increasing production output of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof is desirable, such as in vitro cell free systems for protein production.

In some embodiments, the present invention features a method of increasing expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof by a cell or in vitro expression system, the method comprising contacting the cell or in vitro expression system with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof. In some embodiments, the method is a method of increasing the expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof by a cell comprising contacting the cell with an effective amount of a compound described herein (e.g., the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof). In other embodiments, the method is a method of increasing the expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof by an in vitro protein expression system comprising contacting the in vitro expression system with a compound described herein (e.g. the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof). In some embodiments, contacting the cell or in vitro expression system with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the cell or in vitro expression system by about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%. In some embodiments, contacting the cell or in vitro expression system with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the cell or in vitro expression system by about 1-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 60-fold, about 70-fold, about 80-fold, about 90-fold, about 100-fold, about 200-fold, about 300-fold, about 400-fold, about 500-fold, about 600-fold about 700-fold, about 800-fold, about 900-fold, about 1000-fold, about 10000-fold, about 100000-fold, or about 1000000-fold.

In some embodiments, the present invention features a method of increasing the expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof by a patient cells, the method comprising administering to the patient an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, wherein the patient has been diagnosed with a disease, disorder, or condition disclosed herein and wherein the disease, disorder or condition is characterized by aberrant expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof (e.g., a leukodystrophy, a leukoencephalopathy, a hypomyelinating or demyelinating disease, muscle-wasting disease, or sarcopenia). In some embodiments, administering to the patient in need thereof an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases the expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof by the patients cells about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%, thereby treating the disease, disorder or condition. In some embodiments, administering to the patient in need thereof an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof by the patients cells about 1-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 60-fold, about 70-fold, about 80-fold, about 90-fold, about 100-fold, about 200-fold, about 300-fold, about 400-fold, about 500-fold, about 600-fold about 700-fold, about 800-fold, about 900-fold, about 1000-fold, about 10000-fold, about 100000-fold, or about 1000000-fold, thereby treating the disease, disorder or condition.

In another aspect, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be useful in applications where increasing the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof is desirable.

In some embodiments, the present invention features a method of increasing the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in a cell, the method comprising contacting the cell with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof. In some embodiments, contacting the cell with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the cell by about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%. In some embodiments, contacting the cell with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the cell by about 1-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 60-fold, about 70-fold, about 80-fold, about 90-fold, about 100-fold, about 200-fold, about 300-fold, about 400-fold, about 500-fold, about 600-fold about 700-fold, about 800-fold, about 900-fold, about 1000-fold, about 10000-fold, about 100000-fold, or about 1000000-fold.

In some embodiments, the present invention features a method of increasing the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, wherein the patient has been diagnosed with a disease, disorder, or condition disclosed herein and wherein the disease, disorder or condition is characterized by lowered levels of protein activity. In some embodiments, administering to the patient in need thereof an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the patient by about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%, thereby treating the disease, disorder or condition. In some embodiments, administering to the patient in need thereof an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof increases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the patient by about 1-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 60-fold, about 70-fold, about 80-fold, about 90-fold, about 100-fold, about 200-fold, about 300-fold, about 400-fold, about 500-fold, about 600-fold about 700-fold, about 800-fold, about 900-fold, about 1000-fold, about 10000-fold, about 100000-fold, or about 1000000-fold, thereby treating the disease, disorder or condition.

In some embodiments, the compound of Formula (I) is chemically modified, prior to (ex vivo) or after (in vivo) contacting with the cell or in vitro expression system, forming a biologically active compound that increases the expression and/or activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the cells and/or in vitro expression system. In some embodiments, the compound of Formula (I) is metabolized by the patient forming a biologically active compound that increases the expression and/or activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the patients cells, thereby treating a condition, disease or disorder disclosed herein. In some embodiments, the biologically active compound is the compound of formula (II).

Methods of Decreasing Protein Activity and Production

In another aspect, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be useful in applications where decreasing production output of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof is desirable.

In some embodiments, the present invention features a method of decreasing expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in a cell, the method comprising contacting the cells with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof. In some embodiments, contacting the cells with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof decreases expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the cell by about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

In some embodiments, the present invention features a method of decreasing the expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, wherein the patient has been diagnosed with a disease, disorder, or condition described herein and wherein the disease, disorder or condition is characterized by increased levels of protein production. In some embodiments, administering to the patient in need thereof an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof decreases the expression of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the patient by about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%, thereby treating the disease, disorder or condition.

In another aspect, the compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof may be useful in applications where decreasing the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof is desirable.

In some embodiments, the present invention features a method of decreasing the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in a cell, the method comprising contacting the cell with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof. In some embodiments, contacting the cell with an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof decreases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the cell by about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%, thereby treating the disease, disorder or condition.

In some embodiments, the present invention features a method of decreasing the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in a patient in need thereof, the method comprising administering to the patient an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, wherein the patient has been diagnosed with a disease, disorder, or condition described herein and wherein the disease, disorder or condition is characterized by increased levels of protein activity. In some embodiments, administering to the patient in need thereof an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof decreases the activity of eIF2B, eIF2α, a component of the eIF2 pathway, a component of the ISR pathway or any combination thereof in the patient by about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%, thereby treating the disease, disorder or condition.

In some embodiments, the compound of Formula (I) is chemically modified, prior to (ex vivo) or after (in vivo) contacting with a cell, forming a biologically active compound that decreases the expression and/or activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the cell. In some embodiments, the compound of Formula (I) is metabolized by the patient forming a biologically active compound that decreases the expression and/or activity of eIF2B, eIF2α, a component of the eIF2 pathway, component of the ISR pathway or any combination thereof in the patients cells, thereby treating a condition, disease or disorder disclosed herein. In some embodiments, the biologically active compound is the compound of formula (I).

Combination Therapy

In one aspect, the present invention features a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof as well as a second agent (e.g. a second therapeutic agent). In some embodiments, the pharmaceutical composition includes a second agent (e.g. a second therapeutic agent) in a therapeutically effective amount. In some embodiments, the second agent is an agent for treating cancer, a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, a metabolic disease, or a disease or disorder associated with impaired function of eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway.

The compounds described herein can be used in combination with one another, with other active agents known to be useful in treating cancer, a neurodegenerative disease, an inflammatory disease, a musculoskeletal disease, a metabolic disease, or a disease or disorder associated with impaired function of eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway or with adjunctive agents that may not be effective alone, but may contribute to the efficacy of the active agent.

In some embodiments, co-administration includes administering one active agent within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of a second active agent. Co-administration includes administering two active agents simultaneously, approximately simultaneously (e.g., within about 1, 5, 10, 15, 20, or 30 minutes of each other), or sequentially in any order. In some embodiments, co-administration can be accomplished by co-formulation, i.e., preparing a single pharmaceutical composition including both active agents. In other embodiments, the active agents can be formulated separately. In another embodiment, the active and/or adjunctive agents may be linked or conjugated to one another. In some embodiments, the compounds described herein may be combined with treatments for a cancer, a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, a metabolic disease, or a disease or disorder associated with impaired function of eIF2B, eIF2α, or a component of the eIF2 pathway or ISR pathway.

In embodiments, the second agent is an anti-cancer agent. In embodiments, the second agent is a chemotherapeutic. In embodiments, the second agent is an agent for improving memory. In embodiments, the second agent is an agent for treating a neurodegenerative disease. In embodiments, the second agent is an agent for treating a leukodystrophy. In embodiments, the second agent is an agent for treating vanishing white matter disease. In embodiments, the second agent is an agent for treating childhood ataxia with CNS hypo-myelination. In embodiments, the second agent is an agent for treating an intellectual disability syndrome (e.g., Fragile X syndrome). In embodiments, the second agent is an agent for treating pancreatic cancer. In embodiments, the second agent is an agent for treating breast cancer. In embodiments, the second agent is an agent for treating multiple myeloma. In embodiments, the second agent is an agent for treating myeloma. In embodiments, the second agent is an agent for treating a cancer of a secretory cell. In embodiments, the second agent is an agent for reducing eIF2a phosphorylation. In embodiments, the second agent is an agent for inhibiting a pathway activated by eIF2α phosphorylation. In embodiments, the second agent is an agent for inhibiting a pathway activated by eIF2α. In embodiments, the second agent is an agent for inhibiting the integrated stress response. In embodiments, the second agent is an anti-inflammatory agent. In embodiments, the second agent is an agent for treating postsurgical cognitive dysfunction. In embodiments, the second agent is an agent for treating traumatic brain injury. In embodiments, the second agent is an agent for treating a musculoskeletal disease. In embodiments, the second agent is an agent for treating a metabolic disease. In embodiments, the second agent is an anti-diabetic agent.

Anti-Cancer Agents

“Anti-cancer agent” is used in accordance with its plain ordinary meaning and refers to a composition (e.g. compound, drug, antagonist, inhibitor, modulator) having antineoplastic properties or the ability to inhibit the growth or proliferation of cells. In some embodiments, an anti-cancer agent is a chemotherapeutic. In some embodiments, an anti-cancer agent is an agent identified herein having utility in methods of treating cancer. In some embodiments, an anticancer agent is an agent approved by the FDA or similar regulatory agency of a country other than the USA, for treating cancer. Examples of anti-cancer agents include, but are not limited to, MEK (e.g. MEK1, MEK2, or MEK1 and MEK2) inhibitors (e.g. XL518, CI-1040, PD035901, selumetinib/AZD6244, GSK1120212/trametinib, GDC-0973, ARRY-162, ARRY-300, AZD8330, PD0325901, U0126, PD98059, TAK-733, PD318088, AS703026, BAY 869766), alkylating agents (e.g., cyclophosphamide, ifosfamide, chlorambucil, busulfan, melphalan, mechlorethamine, uramustine, thiotepa, nitrosoureas, nitrogen mustards (e.g., mechloroethamine, cyclophosphamide, chlorambucil, meiphalan), ethylenimine and methylmelamines (e.g., hexamethlymelamine, thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomusitne, semustine, streptozocin), triazenes (decarbazine), anti-metabolites (e.g., 5-azathioprine, leucovorin, capecitabine, fludarabine, gemcitabine, pemetrexed, raltitrexed, folic acid analog (e.g., methotrexate), or pyrimidine analogs (e.g., fluorouracil, floxouridine, Cytarabine), purine analogs (e.g., mercaptopurine, thioguanine, pentostatin), etc.), plant alkaloids (e.g., vincristine, vinblastine, vinorelbine, vindesine, podophyllotoxin, paclitaxel, docetaxel, etc.), topoisomerase inhibitors (e.g., irinotecan, topotecan, amsacrine, etoposide (VP 16), etoposide phosphate, teniposide, etc.), antitumor antibiotics (e.g., doxorubicin, adriamycin, daunorubicin, epirubicin, actinomycin, bleomycin, mitomycin, mitoxantrone, plicamycin, etc.), platinum-based compounds (e.g. cisplatin, oxaloplatin, carboplatin), anthracenedione (e.g., mitoxantrone), substituted urea (e.g., hydroxyurea), methyl hydrazine derivative (e.g., procarbazine), adrenocortical suppressant (e.g., mitotane, aminoglutethimide), epipodophyllotoxins (e.g., etoposide), antibiotics (e.g., daunorubicin, doxorubicin, bleomycin), enzymes (e.g., L-asparaginase), inhibitors of mitogen-activated protein kinase signaling (e.g. U0126, PD98059, PD184352, PD0325901, ARRY-142886, SB239063, SP600125, BAY 43-9006, wortmannin, or LY294002, Syk inhibitors, mTOR inhibitors, antibodies (e.g., rituxan), gossyphol, genasense, polyphenol E, Chlorofusin, all trans-retinoic acid (ATRA), bryostatin, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), 5-aza-2′-deoxycytidine, all trans retinoic acid, doxorubicin, vincristine, etoposide, gemcitabine, imatinib (Gleevec®), geldanamycin, 17-N-Allylamino-17-Demethoxygeldanamycin (17-AAG), flavopiridol, LY294002, bortezomib, trastuzumab, BAY 1 1-7082, PKC412, PD184352, 20-epi-1, 25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; arsenic trioxide; asulacrine; atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine; clomifene analogues; clotrimazole; collismycin A; collismycin B; combretastatin A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone; didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine; 9-dioxamycin; diphenyl spiromustine; docosanol; dolasetron; doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride; estramustine analogue; estrogen agonists; estrogen antagonists; etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride; flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane; fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine; ilomastat; imidazoacridones; imiquimod; immunostimulant peptides; insulin-like growth factor-1 receptor inhibitor; interferon agonists; interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemia inhibiting factor; leukocyte alpha interferon; leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole; linear polyamine analogue; lipophilic disaccharide peptide; lipophilic platinum compounds; lissoclinamide 7; lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysin inhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone; meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone; mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonal antibody, human chorionic gonadotrophin; monophosphoryl lipid A+mycobacterium cell wall sk; mopidamol; multiple drug resistance gene inhibitor; multiple tumor suppressor 1-based therapy; mustard anticancer agent; mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn; 06-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin; osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin; pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A; placetin B; plasminogen activator inhibitor; platinum complex; platinum compounds; platinum-triamine complex; porfimer sodium; porfiromycin; prednisone; propyl bis-acridone; prostaglandin J2; proteasome inhibitors; protein A-based immune modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylerie conjugate; raf antagonists; raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide; rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine; senescence derived inhibitor 1; sense oligonucleotides; signal transduction inhibitors; signal transduction modulators; single chain antigen-binding protein; sizofuran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-cell division inhibitors; stipiamide; stromelysin inhibitors; sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; synthetic glycosaminoglycans; tallimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomerase inhibitors; temoporfin; temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic; thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocene bichloride; topsentin; toremifene; totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase receptor antagonists; vapreotide; variolin B; vector system, erythrocyte gene therapy; velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; zinostatin stimalamer, Adriamycin, Dactinomycin, Bleomycin, Vinblastine, Cisplatin, acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin; altretamine; ambomycin; ametantrone acetate; aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine; dacarbazine; daunorubicin hydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin hydrochloride; estramustine; estramustine phosphate sodium; etanidazole; etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine; fludarabine phosphate; fluorouracil; fluorocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; iimofosine; interleukin II (including recombinant interleukin II, or r1L.sub.2), interferon alfa-2a; interferon alfa-2b; interferon alfa-nl; interferon alfa-n3; interferon beta-1a; interferon gamma-1b; iprop latin; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolide acetate; liarozole hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazoie; nogalamycin; ormaplatin; oxisuran; pegaspargase; peliomycin; pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride; semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifene citrate; trestolone acetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uredepa; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicin hydrochloride, agents that arrest cells in the G2-M phases and/or modulate the formation or stability of microtubules, (e.g. Taxol (i.e. paclitaxel), Taxotere, compounds comprising the taxane skeleton, Erbulozole (i.e. R-55104), Dolastatin 10 (i.e. DLS-10 and NSC-376128), Mivobulin isethionate (i.e. as CI-980), Vincristine, NSC-639829, Discodermolide (i.e. as NVP-XX-A-296), ABT-751 (Abbott, i.e. E-7010), Altorhyrtins (e.g. Altorhyrtin A and Altorhyrtin C), Spongistatins (e.g. Spongistatin 1, Spongistatin 2, Spongistatin 3, Spongistatin 4, Spongistatin 5, Spongistatin 6, Spongistatin 7, Spongistatin 8, and Spongistatin 9), Cemadotin hydrochloride (i.e. LU-103793 and SC-D-669356), Epothilones (e.g. Epothilone A, Epothilone B, Epothilone C (i.e. desoxyepothilone A or dEpoA), Epothilone D (i.e. KOS-862, dEpoB, and desoxyepothilone B), Epothilone E, Epothilone F, Epothilone B N-oxide, Epothilone A N-oxide, 16-aza-epothilone B, 21-aminoepothilone B (i.e. BMS-310705), 21-hydroxyepothilone D (i.e. Desoxyepothilone F and dEpoF), 26-fluoroepothilone, Auristatin PE (i.e. NSC-654663), Soblidotin (i.e. TZT-1027), LS-4559-P (Pharmacia, i.e. LS-4577), LS-4578 (Pharmacia, i.e. LS-477-P), LS-4477 (Pharmacia), LS-4559 (Pharmacia), RPR-1 12378 (Aventis), Vincristine sulfate, DZ-3358 (Daiichi), FR-182877 (Fujisawa, i.e. WS-9885B), GS-164 (Takeda), GS-198 (Takeda), KAR-2 (Hungarian Academy of Sciences), BSF-223651 (BASF, i.e. ILX-651 and LU-223651), SAH-49960 (Lilly/Novartis), SDZ-268970 (Lilly/Novartis), AM-97 (Armad/Kyowa Hakko), AM-132 (Armad), AM-138 (Armad/Kyowa Hakko), IDN-5005 (Indena), Cryptophycin 52 (i.e. LY-355703), AC-7739 (Ajinomoto, i.e. AVE-8063A and CS-39.HCl), AC-7700 (Ajinomoto, i.e. AVE-8062, AVE-8062A, CS-39-L-Ser.HCl, and RPR-258062A), Vitilevuamide, Tubulysin A, Canadensol, Centaureidin (i.e. NSC-106969), T-138067 (Tularik, i.e. T-67, TL-138067 and TI-138067), COBRA-1 (Parker Hughes Institute, i.e. DDE-261 and WHI-261), H10 (Kansas State University), H16 (Kansas State University), Oncocidin A 1 (i.e. BTO-956 and DIME), DDE-313 (Parker Hughes Institute), Fijianolide B, Laulimalide, SPA-2 (Parker Hughes Institute), SPA-1 (Parker Hughes Institute, i.e. SPIKET-P), 3-IAABU (Cytoskeleton/Mt. Sinai School of Medicine, i.e. MF-569), Narcosine (also known as NSC-5366), Nascapine, D-24851 (Asta Medica), A-105972 (Abbott), Hemiasterlin, 3-BAABU (Cytoskeleton/Mt. Sinai School of Medicine, i.e. MF-191), TMPN (Arizona State University), Vanadocene acetylacetonate, T-138026 (Tularik), Monsatrol, Inanocine (i.e. NSC-698666), 3-IAABE (Cytoskeleton/Mt. Sinai School of Medicine), A-204197 (Abbott), T-607 (Tularik, i.e. T-900607), RPR-115781 (Aventis), Eleutherobins (such as Desmethyleleutherobin, Desaetyleleutherobin, lsoeleutherobin A, and Z-Eleutherobin), Caribaeoside, Caribaeolin, Halichondrin B, D-64131 (Asta Medica), D-68144 (Asta Medica), Diazonamide A, A-293620 (Abbott), NPI-2350 (Nereus), Taccalonolide A, TUB-245 (Aventis), A-259754 (Abbott), Diozostatin, (−)-Phenylahistin (i.e. NSCL-96F037), D-68838 (Asta Medica), D-68836 (Asta Medica), Myoseverin B, D-43411 (Zentaris, i.e. D-81862), A-289099 (Abbott), A-318315 (Abbott), HTI-286 (i.e. SPA-110, trifluoroacetate salt) (Wyeth), D-82317 (Zentaris), D-82318 (Zentaris), SC-12983 (NCO, Resverastatin phosphate sodium, BPR-OY-007 (National Health Research Institutes), and SSR-25041 1 (Sanofi), steroids (e.g., dexamethasone), finasteride, aromatase inhibitors, gonadotropin-releasing hormone agonists (GnRH) such as goserelin or leuprolide, adrenocorticosteroids (e.g., prednisone), progestins (e.g., hydroxyprogesterone caproate, megestrol acetate, medroxyprogesterone acetate), estrogens (e.g., diethlystilbestrol, ethinyl estradiol), antiestrogen (e.g., tamoxifen), androgens (e.g., testosterone propionate, fluoxymesterone), antiandrogen (e.g., flutamide), immunostimulants (e.g., Bacillus Calmette-Guerin (BCG), levamisole, interleukin-2, alpha-interferon, etc.), monoclonal antibodies (e.g., anti-CD20, anti-HER2, anti-CD52, anti-HLA-DR, and anti-VEGF monoclonal antibodies), immunotoxins (e.g., anti-CD33 monoclonal antibody-calicheamicin conjugate, anti-CD22 monoclonal antibody-pseudomonas exotoxin conjugate, etc.), radioimmunotherapy (e.g., anti-CD20 monoclonal antibody conjugated to ^(U 1)ln, ⁹⁰Y, or ¹³¹I, etc.), triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, topotecan, itraconazole, vindesine, cerivastatin, vincristine, deoxyadenosine, sertraline, pitavastatin, irinotecan, clofazimine, 5-nonyloxytryptamine, vemurafenib, dabrafenib, erlotinib, gefitinib, EGFR inhibitors, epidermal growth factor receptor (EGFR)-targeted therapy or therapeutic (e.g. gefitinib (Iressa™), erlotinib (Tarceva™), cetuximab (Erbitux™), lapatinib (Tykerb™), panitumumab (Vectibix™) vandetanib (Caprelsa™), afatinib/BIBW2992, CI-1033/canertinib, neratinib/HKI-272, CP-724714, TAK-285, AST-1306, ARRY334543, ARRY-380, AG-1478, dacomitinib/PF299804, OSI-420/desmethyl erlotinib, AZD8931, AEE788, pelitinib/EKB-569, CUDC-101, WZ8040, WZ4002, WZ3146, AG-490, XL647, PD153035, BMS-599626), sorafenib, imatinib, sunitinib, dasatinib, or the like.

“Chemotherapeutic” or “chemotherapeutic agent” is used in accordance with its plain ordinary meaning and refers to a chemical composition or compound having antineoplastic properties or the ability to inhibit the growth or proliferation of cells.

Additionally, the compounds described herein can be co-administered with conventional immunotherapeutic agents including, but not limited to, immunostimulants (e.g., Bacillus Calmette-Guerin (BCG), levamisole, interleukin-2, alpha-interferon, etc.), monoclonal antibodies (e.g., anti-CD20, anti-HER2, anti-CD52, anti-HLA-DR, and anti-VEGF monoclonal antibodies), immunotoxins (e.g., anti-CD33 monoclonal antibody-calicheamicin conjugate, anti-CD22 monoclonal antibody-pseudomonas exotoxin conjugate, etc.), and radioimmunotherapy (e.g., anti-CD20 monoclonal antibody conjugated to ^(m)In, ⁹⁰Y, or ¹³¹I, etc.).

In a further embodiment, the compounds described herein can be co-administered with conventional radiotherapeutic agents including, but not limited to, radionuclides such as ⁴⁷Sc, ⁶⁴Cu, ⁶⁷Cu, ⁸⁹Sr, ⁸⁶Y, ⁸⁷Y, ⁹⁰Y, ¹⁰⁵Rh, mAg, mIn, ¹¹⁷mSn, ¹⁴⁹Pm, ¹⁵³Sm, ¹⁶⁶Ho, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At, and ²¹²Bi, optionally conjugated to antibodies directed against tumor antigens.

Additional Agents

In some embodiments, the second agent for use in combination with a compound (e.g., a compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof), or composition thereof described herein is an agent for use in treating a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, or a metabolic disease. In some embodiments, a second agent for use in combination with a compound (e.g., a compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof) or composition thereof described herein is an agent approved by the FDA or similar regulatory agency of a country other than the USA, for treating a disease, disorder, or condition described herein.

In some embodiments, a second agent for use in treating a neurodegenerative disease, a leukodystrophy, an inflammatory disease, a musculoskeletal disease, or a metabolic disease includes, but is not limited to, an anti-psychotic drug, anti-depressive drug, anti-anxiety drug, analgesic, a stimulant, a sedative, a pain reliever, an anti-inflammatory agent, a benzodiazepine, a cholinesterase inhibitor, a non-steroidal anti-inflammatory drug (NSAID), a corticosteroid, a MAO inhibitor, a beta-blocker, a calcium channel blocker, an antacid, or other agent. Exemplary second agents may include donepezil, galantamine, rivastigmine, memantine, levodopa, dopamine, pramipexole, ropinirole, rotigotine, doxapram, oxazepam, quetiapine, selegiline, rasagiline, entacapone, benztropine, trihexyphenidyl, riluzole, diazepam, chlorodiazepoxide, lorazepam, alprazolam, buspirone, gepirone, ispapirone, hydroxyzine, propranolol, hydroxyzine, midazolam, trifluoperazine, methylphenidate, atomoxetine, methylphenidate, pemoline, perphenazine, divalproex, valproic acid, sertraline, fluoxetine, citalopram, escitalopram, paroxetine, fluvoxamine, trazodone, desvenlafaxine, duloxetine, venlafaxine, amitriptyline, amoxapine, clomipramine, desipramine, imipramine, nortriptyline, protriptyline, trimipramine, maprotiline, bupropion, nefazodone, vortioxetine, lithium, clozapine, fluphenazine, haloperidol, paliperidone, loxapine, thiothixene, pimozide, thioridazine, risperidone, aspirin, ibuprofen, naproxen, acetaminophen, azathioprine, methotrexate, mycophenolic acid, leflunomide, dibenzoylmethane, cilostazol, pentoxifylline, duloxetine, a cannabinoid (e.g., nabilone), simethicone, magaldrate, aluminum salts, calcium salts, sodium salts, magnesium salts, alginic acid, acarbose, albiglutide, alogliptin, metformin, insulin, lisinopril, atenolol, atorvastatin, fluvastatin, lovastatin, pitavastatin, simvastatin, rosuvastatin, and the like.

Naturally derived agents or supplements may also be used in conjunction with a compound of Formula (I) or a pharmaceutically acceptable salt, co-crystal, solvate, hydrate, tautomer, ester, N-oxide or stereoisomer thereof, or a composition thereof to treat a neurodegenerative disease, an inflammatory disease, a musculoskeletal disease, or a metabolic disease. Exemplary naturally derived agents or supplements include omega-3 fatty acids, carnitine, citicoline, curcumin, gingko, vitamin E, vitamin B (e.g., vitamin B5, vitamin B6, or vitamin B12), huperzine A, phosphatidylserine, rosemary, caffeine, melatonin, chamomile, St. John's wort, tryptophan, and the like.

EXAMPLES

In order that the invention described herein may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting their scope.

Synthetic Protocols

The compounds provided herein can be prepared from readily available starting materials using modifications to the specific synthesis protocols set forth below that would be well known to those of skill in the art. It will be appreciated that where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by those skilled in the art by routine optimization procedures. General scheme relating to methods of making exemplary compounds of the invention are additionally described in the section entitled Methods of Making Compounds.

Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions for protection and deprotection are well known in the art. For example, numerous protecting groups, and their introduction and removal, are described in Greene et al., Protecting Groups in Organic Synthesis, Second Edition, Wiley, New York, 1991, and references cited therein.

Abbreviations

APCI for atmospheric pressure chemical ionization; Boc for tert-butoxycarbonyl; t-Bu for tert-butyl; DCI for desorption chemical ionization; DMSO for dimethyl sulfoxide; ESI for electrospray ionization; HATU for 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate; HPLC for high performance liquid chromatography; LC for liquid chromatography; LC/MS for liquid chromatography/mass spectrometry; MS for mass spectrum; NMR for nuclear magnetic resonance; psi for pounds per square inch; SFC for supercritical fluid chromatography; and UV for ultraviolet.

Example 1: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl acetate (Compound 100) Example 1A: ethyl 1,4-dioxaspiro[4.5]decane-8-carboxylate

A mixture of ethyl 4-oxocyclohexanecarboxylate (11.70 mL, 73.4 mmol), ethane-1,2-diol (12.29 mL, 220 mmol), and p-toluenesulfonic acid monohydrate (1.397 g, 7.34 mmol) in toluene (200 mL) was stirred at 120° C. with a Dean-Stark trap apparatus for 180 minutes. The reaction mixture was neutralized with N-ethyl-N-isopropylpropan-2-amine and then concentrated under reduced pressure. The residue was purified on silica gel (0-30% ethyl acetate in heptane) to give 12.77 g of the title compound as a clear oil. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 4.01 (q, J=7.1 Hz, 2H), 3.81 (s, 4H), 2.32 (tt, J=10.4, 3.8 Hz, 1H), 1.83-1.71 (m, 2H), 1.66-1.57 (m, 1H), 1.62-1.38 (m, 5H), 1.13 (t, J=7.1 Hz, 3H).

Example 1B: ethyl 8-acetyl-1,4-dioxaspiro[4.5]decane-8-carboxylate

To a solution of diisopropylamine (5.19 mL, 36.4 mmol) in tetrahydrofuran (25 mL) at 0° C. was added n-butyllithium slowly with internal temperature being maintained below 5° C. After stirring for 30 minutes, the solution was cooled to −78° C. under nitrogen, and a solution of Example 1A (6.0 g, 28.0 mmol) in tetrahydrofuran (3 mL) was added slowly, and the resultant mixture was stirred for 30 minutes at the same temperature. Then acetyl chloride (2.59 mL, 36.4 mmol) was added slowly to maintain the temperature below −60° C., and the mixture was stirred at −70° C. for 2 hours. The reaction was quenched with saturated, aqueous NH₄Cl solution, and the aqueous phase was extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified on silica gel (0-70% ethyl acetate in heptane) to give 6.78 g of the title compound as a clear oil. ¹H NMR (500 MHz, DMSO-d₆) δ ppm 4.19-4.11 (m, 2H), 3.85 (s, 4H), 2.13 (s, 3H), 2.10-2.01 (m, 2H), 1.90 (ddd, J=13.9, 9.6, 4.6 Hz, 2H), 1.54 (th, J=13.6, 4.7 Hz, 4H), 1.18 (dd, J=7.6, 6.5 Hz, 3H).

Example 1C: ethyl 1-acetyl-4-oxocyclohexane-1-carboxylate

A mixture of Example 1B (6.5 g, 25.4 mmol) and HCl (21.13 mL, 127 mmol) in acetone (60 mL) was stirred at ambient temperature overnight. Volatiles were removed under reduced pressure, and the residue was partitioned between water and dichloromethane. The organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure to give 5.46 g of the title compound as a clear oil which was used without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 4.16 (q, J=7.1 Hz, 2H), 2.35-2.07 (m, 8H), 2.17 (s, 3H), 1.17 (t, J=7.1 Hz, 3H).

Example 1D: ethyl 4-(benzylamino)-2-oxobicyclo[2.2.2]octane-1-carboxylate

A mixture of Example 1C (9.7 g, 45.7 mmol), benzylamine (14.98 mL, 137 mmol), and p-toluenesulfonic acid monohydrate (0.087 g, 0.457 mmol) in toluene (100 mL) was stirred at reflux with a Dean-Stark trap apparatus overnight. The mixture was concentrated under reduced pressure, and the residue was stirred with a mixture of ethyl acetate (50 mL) and 3 N HCl (100 mL) for 30 minutes. The precipitate was collected by filtration, washed with a mixture of ethyl acetate/heptane and air-dried to give 11.3 g of title compound as an HCl salt. The filtrate was neutralized with 6 N NaOH and extracted with ethyl acetate (100 mL×2). The organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered under reduced pressure. The residue was purified on silica gel (0-70% ethyl acetate in heptane) to give another 0.77 g of the title compound as yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.73 (t, J 6.2 Hz, 2H), 7.87-7.12 (m, 5H), 4.09 (m, 4H), 2.88 (s, 2H), 2.08 (dt, J=20.7, 13.4 Hz, 6H), 1.16 (t, J=7.1 Hz, 3H); MS (ESI⁺) m/z 302.1 (M+H)⁺.

Example 1E: ethyl 4-amino-2-oxobicyclo[2.2.2]octane-1-carboxylate

To a mixture of Example 1D (11.2 g, 33.2 mmol) in tetrahydrofuran (110 mL) in a 50 mL pressure bottle was added 20% Pd(OH)₂/C, wet (2.2 g, 1.6 mmol), and the reaction was shaken at 50° C. under 50 psi of hydrogen for 22 hours. The reaction mixture was cooled to ambient temperature, and then the solids were removed by filtration and washed with methanol (1 L). The filtrate and wash were concentrated under reduced pressure to give 7.9 g of the title compound as a hydrochloride salt. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.46 (s, 3H), 4.07 (q, J=7.1 Hz, 2H), 2.62 (s, 2H), 2.17-2.05 (m, 2H), 2.04-1.78 (m, 6H), 1.14 (t, J=7.1 Hz, 3H).

Example 1F: ethyl 4-[2-(4-chloro-3-fluorophenoxy)acetamido]-2-oxobicyclo[2.2.2]octane-1-carboxylate

To a suspension of Example 1E (7.8 g, 31.5 mmol), N-ethyl-N-isopropylpropan-2-amine (22.00 mL, 126 mmol) and 2-(4-chloro-3-fluorophenoxy)acetic acid (7.41 g, 36.2 mmol) in N,N-dimethylformamide (200 mL), was added 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate(V) (14.97 g, 39.4 mmol), and the resulting brown solution was stirred at ambient temperature for 16 hours. Water was added, and the mixture was stirred for 15 minutes. The precipitate was collected by filtration, washed with water, and air-dried to give 12.1 g of the title compound as an off-white solid. ¹H NMR (400 MHz, DMSO-d₆) (5 ppm 7.87 (s, 1H), 7.45 (t, J=8.9 Hz, 1H), 7.00 (dd, J=11.4, 2.9 Hz, 1H), 6.79 (ddd, J=8.9, 2.9, 1.2 Hz, 1H), 4.45 (s, 2H), 4.06 (q, J=7.1 Hz, 2H), 2.73 (s, 2H), 2.07 (m, 1H), 2.01-1.84 (m, 6H), 1.14 (t, J=7.1 Hz, 3H); MS (ESI⁺) m/z 398.0 (M+H)⁺.

Example 1G: 4-[2-(4-chloro-3-fluorophenoxy)acetamido]-2-oxobicyclo[2.2.2]octane-1-carboxylic Acid

A suspension of Example 1F (11.37 g, 28.6 mmol) and sodium hydroxide (7.15 mL, 57.2 mmol, 8 M solution) in methanol (100 mL) was stirred at ambient temperature for 16 hours. Volatiles were removed under reduced pressure, and the residue was acidified with 1 N HCl. The precipitate was collected by filtration and dried in a vacuum oven to give 9.9 g of the title compound as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.49 (s, 1H), 7.86 (s, 1H), 7.45 (t, J=8.9 Hz, 1H), 7.00 (dd, J=11.4, 2.9 Hz, 1H), 6.83-6.74 (m, 1H), 4.45 (s, 2H), 2.71 (s, 2H), 2.01-1.81 (m, 7H); MS (ESI) m/z 368.1 (M−H)⁻.

Example 1H: N-(4-amino-3-oxobicyclo[2.2.2]octan-1-yl)-2-(4-chloro-3-fluorophenoxy)acetamide

A mixture of Example 1G (3.24 g, 8.76 mmol), diphenylphosphoryl azide (2.84 mL, 13.14 mmol), and triethylamine (3.66 mL, 26.3 mmol) in toluene (100 mL) was heated at 110° C. for 2 hours. The solution was cooled to ambient temperature and poured into 150 mL of 3 N HCl solution. The mixture was stirred for 16 hours to give a suspension. The precipitate was filtered, washed with ethyl acetate, and air-dried to give the title compound (1.63 g) as an HCl salt as a white solid. The filtrate was then basified with solid sodium bicarbonate and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified on silica gel (0-10% methanol/dichloromethane) to give the title compound (0.6 g) as the free base. ¹H NMR (400 MHz, DMSO-d₆) (5 ppm 8.49 (s, 3H), 8.08 (s, 1H), 7.45 (t, J=8.9 Hz, 1H), 7.01 (dd, J=11.4, 2.8 Hz, 1H), 6.79 (ddd, J=9.0, 2.9, 1.2 Hz, 1H), 4.48 (s, 2H), 2.90 (s, 2H), 2.12-1.79 (m, 8H).

Example 1I: N-(4-amino-3-hydroxybicyclo[2.2.2]octan-1-yl)-2-(4-chloro-3-fluorophenoxy)acetamide hydrochloride

A mixture of Example 1H (2.5 g, 6.63 mmol) and sodium borohydride (1.254 g, 33.1 mmol) in a 1:1 mixture of methanol/dichloromethane (50 mL) was stirred for 24 hours. Volatiles were removed under reduced pressure, and the residue was partitioned between water and dichloromethane. The organic fraction was separated, dried (MgSO₄), and concentrated. The residue was then treated with 4 N HCl in dioxane. The suspension was sonicated and concentrated. The residue was dried under vacuum to give 2.82 g of the title compound as a light yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.97 (s, 3H), 7.72 (s, 1H), 7.40 (t, J=8.9 Hz, 1H), 6.95 (dd, J=11.4, 2.8 Hz, 1H), 6.74 (ddd, J=9.0, 2.9, 1.1 Hz, 1H), 5.64 (s, 1H), 4.41 (s, 2H), 3.83 (d, J=9.1 Hz, 1H), 2.24 (td, J=10.8, 9.9, 5.3 Hz, 1H), 1.96-1.51 (m, 9H); MS (ESI⁺) m/z 343.0 (M+H)⁺.

Example 1J: N,N′-(2-oxobicyclo[2.2.2]octane-1,4-diyl)bis[2-(4-chloro-3-fluorophenoxy)acetamide]

A mixture of Example 1H (0.5 g, 1.325 mmol), 2-(4-chloro-3-fluorophenoxy)acetic acid (0.339 g, 1.657 mmol) and N-ethyl-N-isopropylpropan-2-amine (1.157 mL, 6.63 mmol) in N,N-dimethylformamide (20 mL) was treated with 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate(V) (0.756 g, 1.988 mmol), and the reaction mixture was stirred at ambient temperature for 30 minutes to see a complete conversion. Water was added, and the resultant mixture was stirred for 15 minutes. The precipitate was collected by filtration, washed with water, and dried in vacuum oven at 50° C. for 2 hours to give 0.64 g of the title compound as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.88 (s, 1H), 7.67 (s, 1H), 7.45 (td, J=8.9, 2.4 Hz, 2H), 7.03 (ddd, J=15.0, 11.4, 2.8 Hz, 2H), 6.80 (dddd, J=10.3, 8.9, 2.9, 1.2 Hz, 2H), 4.53 (s, 2H), 4.45 (s, 2H), 2.83 (s, 2H), 2.45-2.33 (m, 2H), 2.10-1.90 (m, 4H), 1.81 (td, J=11.6, 6.3 Hz, 2H); MS (EST⁺) m/z 527.0 (M+H)⁺.

Example 1K: N,N′-(2-hydroxybicyclo[2.2.2]octane-1,4-diyl)bis[2-(4-chloro-3-fluorophenoxy)acetamide]

To a solution of Example 1J (0.63 g, 1.195 mmol) in dichloromethane (10 mL) and methanol (10 mL), sodium borohydride (0.226 g, 5.97 mmol) was added portionwise, and the mixture was stirred at ambient temperature for 4 hours. Volatiles were removed, and the residue was triturated with dichloromethane/methanol to give 0.32 g of the title compound as a white solid. The filtrate was concentrated, and the residue was purified on silica gel (10-100% ethyl acetate in heptane) to give 0.21 g of the title compound. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.49-7.39 (m, 3H), 7.22 (s, 1H), 7.00 (ddd, J=12.4, 11.4, 2.8 Hz, 2H), 6.78 (tdd, J=9.1, 2.9, 1.2 Hz, 2H), 5.04 (s, 1H), 4.41 (d, J=13.3 Hz, 4H), 4.00 (dd, J=9.6, 3.1 Hz, 1H), 2.23 (ddd, J=12.1, 9.4, 2.2 Hz, 1H), 2.09-1.97 (m, 1H), 1.93-1.80 (m, 2H), 1.84-1.68 (m, 6H); MS (ESI⁺) m/z 529.1 (M+H)⁺.

Example 1L: N—N′-[(2S)-2-hydroxybicyclo[2.2.2]octane-1,4-diyl]bis[2-(4-chloro-3-fluorophenoxy)acetamide]

The title compound was isolated by chiral preparative SFC (Supercritical Fluid Chromatography) of Example 1K as the second peak eluted off the column. Preparative SFC was performed on a THAR/Waters SFC 80 system running under SuperChrom™ software control. The preparative SFC system was equipped with an 8-way preparative column switcher, CO₂ pump, modifier pump, automated back pressure regulator (ABPR), UV detector, and 6-position fraction collector. The mobile phase was comprised of supercritical CO₂ supplied by a Dewar of bone-dry non-certified CO₂ pressurized to 350 psi with a modifier of methanol at a flow rate of 70 g/minute. The column was at ambient temperature, and the backpressure regulator was set to maintain 100 bar. The sample was dissolved in a mixture of methanol/dichloromethane (1:1) at a concentration of 10 mg/mL. The sample was loaded into the modifier stream in 1 mL (10 mg) injections. The mobile phase was held isocratically at 30% methanol:CO₂. Fraction collection was time triggered. The instrument was fitted with a Chiralpak® AD-H column with dimensions 21 mm i.d.×250 mm length with 5 μm particles. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.49-7.39 (m, 3H), 7.23 (s, 1H), 7.00 (ddd, J=12.4, 11.4, 2.9 Hz, 2H), 6.78 (dddd, J=9.0, 8.0, 2.9, 1.2 Hz, 2H), 5.05 (s, 1H), 4.41 (d, J=13.5 Hz, 4H), 4.00 (dd, J=9.4, 3.0 Hz, 1H), 2.23 (ddd, J=12.3, 9.4, 2.3 Hz, 1H), 2.03 (ddd, J=12.3, 10.5, 4.7 Hz, 1H), 1.89 (d, J=10.7 Hz, 2H), 1.87-1.76 (m, 1H), 1.74 (ddd, J=12.6, 6.7, 2.4 Hz, 5H); MS (EST⁺) m/z 529.1 (M+H)⁺.

Example 1M: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Acetate

To a suspension of the product of Example 1K (7.80 g, 14.73 mmol), N,N-dimethylpyridin-4-amine (1.88 g, 15.4 mmol), and triethylamine (2.70 mL, 19.4 mmol) in CH₂Cl₂ (80 mL) at 0° C. was added acetyl chloride (1.35 mL, 19 mmol). The ice-water bath was removed, and the mixture was stirred for 30 minutes, and concentrated under reduced pressure. The residue was diluted with ethyl acetate. The mixture was washed with 1 N HCl (40 mL), water (25 mL), saturated, aqueous NaHCO₃ (25 mL), and brine (25 mL). The organic fraction was dried over anhydrous Na₂SO₄, adsorbed onto silica gel, and purified on silica gel (25-30% ethyl acetate/dichloromethane) to give 7.78 g of racemic material as a white solid. The enantiomers were separated by chiral preparative SFC (Supercritical Fluid Chromatography). Preparative SFC was performed on a THAR/Waters SFC 80 system running under SuperChrom™ software control. The preparative SFC system was equipped with an 8-way preparative column switcher, CO₂ pump, modifier pump, automated back pressure regulator (ABPR), UV detector, and 6-position fraction collector. The mobile phase was comprised of supercritical CO₂ supplied by a Dewar of bone-dry non-certified CO₂ pressurized to 350 psi with a modifier of methanol at a flow rate of 80 mL/minute. The column was at ambient temperature, and the backpressure regulator was set to maintain 100 bar. The sample was dissolved in a mixture of methanol/dichloromethane (1:1) at a concentration of 10 mg/mL. The sample was loaded into the modifier stream in 1 mL (10 mg) injections. The mobile phase was held isocratically at 40% methanol:CO₂. Fraction collection was time triggered. The instrument was fitted with a Chiralpak® AD-H column with dimensions 21 mm i.d.×250 mm length with 5 μm particles.

The second eluting compound from this chiral separation was the title compound (2.97 g, 5.19 mmol, 35% yield). ¹H NMR (501 MHz, DMSO-d₆) δ ppm 7.60 (s, 1H), 7.55 (s, 1H), 7.47 (td, J=8.9, 2.0 Hz, 2H), 7.00 (ddd, J=16.6, 11.4, 2.9 Hz, 2H), 6.79 (dddd, J=8.6, 7.1, 2.9, 1.1 Hz, 2H), 5.28 (dd, J=9.5, 2.4 Hz, 1H), 4.48-4.39 (m, 4H), 2.39 (ddd, J=14.1, 9.4, 2.9 Hz, 1H), 2.21 (dd, J=11.6, 7.6 Hz, 1H), 2.02 (td, J=8.4, 2.8 Hz, 1H), 1.98 (s, 3H), 1.93 (dd, J=9.1, 6.9 Hz, 2H), 1.89-1.76 (m, 5H); MS (ESI⁺) m/z 570.7 (M+H)⁺.

Example 2: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl dihydrogen Phosphate (Compound 101) Example 2A: (S)-1,4-bis(2-(4-chloro-3-fluorophenoxy)acetamido)bicyclo[2.2.2]octan-2-yl di-tert-butyl Phosphate

To a solution of the product of Example 1L (0.200 g, 0.378 mmol) in 1N,N-dimethylformamide (1.1 mL) was added a 0.45 M acetonitrile solution of 1H-tetrazole (2.1 mL, 0.945 mmol) followed by di-tert-butyl diethylphosphoramidite (0.21 mL, 0.76 mmol), and the mixture was stirred at 50° C. for 1 hour and then was allowed to cool to room temperature. Hydrogen peroxide (0.39 mL, 3.82 mmol) was added to the reaction at room temperature which was then stirred for 2.5 hours. The reaction mixture was diluted with ethyl acetate, washed with saturated, aqueous NaHCO₃ and brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified on silica gel (3% ethanol/dichloromethane). The still impure material was repurified on silica gel (40% ethyl acetate/dichloromethane) to give the title compound (0.124 g, 0.172 mmol, 46% yield), which was carried on without further purification. LC/MS (APCI⁺) m/z 721 (M+H)⁺.

Example 2B: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl dihydrogen Phosphate

A solution of Example 2A (0.1244 g, 0.172 mmol) in CH₂Cl₂ (0.60 mL) and 2,2,2-trifluoroacetic acid (0.20 mL, 2.61 mmol) was stirred for 16 hours and then was concentrated under reduced pressure. The residue was purified by reverse phase chromatography (Phenomenex® Luna® 250×30 mm 10 μm C18 column, 60 mL/min, 15-95% gradient of acetonitrile in 0.1% aqueous trifluoroacetic acid) to give the title compound (0.035 g, 0.058 mmol, 34% yield). ¹H NMR (400 MHz, DMSO-d₆) (5 ppm 7.76 (s, 1H), 7.62 (s, 1H), 7.47 (td, J=8.9, 4.2 Hz, 2H), 7.07 (ddd, J=33.3, 11.4, 2.9 Hz, 2H), 6.85 (dddd, J=31.0, 9.0, 2.9, 1.2 Hz, 2H), 4.59 (t, J=9.0 Hz, 1H), 4.44 (s, 2H), 4.41 (dd, J=3.3 Hz, 2H), 2.45-2.28 (m, 2H), 2.18 (td, J=12.8, 12.3, 5.7 Hz, 1H), 2.11-1.97 (m, 2H), 1.79 (dt, J=14.5, 9.6 Hz, 5H); MS (ESI) m/z 607 (M−H)⁻.

Example 3: (2S)-2-amino-4-({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)-4-oxobutanoic Acid (Compound 102)

A suspension of the product of Example 1L (30 mg, 0.057 mmol) in N,N-dimethylformamide (1 mL) and N,N-diisopropylethylamine (0.025 mL, 0.142 mmol) was treated with 1-[bis(dimethylamino)methylene]3-oxid hexafluorophosphate (24 mg, 0.062 mmol) and with (5)-2-((tert-butoxycarbonyl)amino)succinic acid (20 mg, 0.086 mmol). The reaction mixture was stirred at 25° C. for 20 hours and then was concentrated under reduced pressure. The residue was diluted with trifluoroacetic acid (0.2 mL) and purified by HPLC (Phenomenex® Luna® C18(2) 5 μm 100 Å AXIA™ column 250 mm×21.2 mm, flow rate 25 mL/minute, 0-60% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid in water)) to give the title compound as the trifluoroacetic acid salt (24 mg, 0.03 mmol, 53% yield). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.32 (d, J=22.4 Hz, 3H), 7.66 (d, J=14.0 Hz, 1H), 7.55-7.41 (m, 3H), 7.07-6.95 (m, 2H), 6.84-6.74 (m, 2H), 5.55-5.34 (m, 1H), 4.50-4.38 (m, 4H), 4.38-4.21 (m, 1H), 2.96-2.83 (m. 2H), 2.48-2.40 (m, 1H), 2.27 (d, J=12.5 Hz, 1H), 2.13-1.68 (m, 8H); MS (ESI⁺) m/z 645 (M+H)⁺.

Example 4: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (2-methoxyethoxy)acetate (Compound 103)

A suspension of the product of Example 1L (30 mg, 0.057 mmol) in N,N-dimethylformamide (1 mL) and N,N′-diisopropylethylamine (0.02.5 mL, 0.142 mmol) was treated with 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (24 mg, 0.062 mmol) and 2-(2-methoxyethoxy) acetic acid (11 mg, 0,082 mmol). The reaction mixture was stirred at 25° C. for 20 hours and then was concentrated under reduced pressure. The residue was purified on HPLC (Phenomenex® Luna® C18(2) 5 μm 100 Å AXIA™ column 250 mm×21.2 mm, flow rate 25 mL/minute, 10-80% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid in water)) to give the title compound (18 mg, 0.027 mmol, 49% yield). ^(r)H NMR (500 MHz, DMSO-d₆) δ ppm 7.61 (s, 1H), 7.55 (s, 1H), 7.47 (td, J=8.8, 2.2 Hz, 2H), 7.01 (ddd, J=11.0, 7.8, 2.9 Hz, 2H), 6.79 (id, J=9.4, 3.0 Hz, 2H), 5.43-5.36 (m, 1H), 4.48-4.37 (m, 4H), 4.09 (d, J=3.2, Hz, 2H), 3.64-3.54 (m, 2H), 3.44 (t, J=4.7 Hz, 2H), 3.23 (s, 3H), 2.43 (dt, J=12.8, 5.7 Hz, 1H), 2.23 (d, J=12.5 Hz, 1H), 2.00 (dd, J=13.1, 5.9 Hz, 1H), 1.97-1.84 (m, 6H), 1.88-1.79 (m, 1H); MS (ESI⁺) m/z 646 (M+H)⁺.

Example 5: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Methoxyacetate (Compound 104)

A suspension of the product of Example 1L (30 mg, 0.057 mmol) in N,N-dimethylformamide (1 mL) and NAT-diisopropylethylamine (0.025 mL, 0.142 mmol) was treated, with 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (24 mg, 0.062 mmol) and 2-methoxy acetic acid (8 mg, 0.089 mmol). The reaction mixture was stirred at 25° C. for 20 hours and then was concentrated under reduced pressure. The residue was purified on HPLC (Phenomenex® Luna® C18(2) 5 μm 100 Å AXIA™ column 250 mm×21.2 mm, flow rate 25 mL/minute, 10-80% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid in water)) to give the title compound (17 mg, 0.028 mmol, 50% yield). NMR (501 MHz, DMSO-d₆) δ ppm 7.62 (s, 1H), 7.58 (s, 1H), 7.47 (Id, 8.9, 3.8 Hz, 2H), 7.01 (td, 11.2, 2.9 Hz, 2H), 6.79 (tdd, J=10.0, 2.8, 1.2 Hz, 2H), 5.40 (dd, 9.6, 2.2 Hz, 1H), 4.48-4.37 (m, 4H), 4.06-3.95 (m, 2H), 3.31 (s, 3H), 2.47-2.38 (m, 1H), 2.23 (d, J=13.3 Hz, 1H), 2.04-1.96 (m, 1H), 1.96-1.90 (m, 2H), 1.85 (ddd, J=24.9, 11.7, 5.7 Hz, 5H); MS (ESI⁺) m/z 602 (M+H)⁺.

Example 6: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Ethoxyacetate (Compound 105)

A suspension of product of Example 1L (30 mg, 0.057 mmol) in N,N-dimethylformamide (1 mL) and NAT-diisopropylethylamine (0.025 mL, 0.142 mmol) was treated with 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (24 mg, 0.062 mmol) and 2-ethoxy acetic acid (9 mg, 0.086 mmol). The reaction mixture was stirred at 25° C. for 20 hours and then was concentrated under reduced pressure. The residue was purified on HPLC (Phenomenex® Luna® C18(2) 5 μm 100 Å AXIA™ column 250 mm×21.2 mm, flow rate 25 mL/minute, 10-80% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid in water)) to give the title compound (16 mg, 0.026 mmol 46% yield). ¹H NMR (501 MHz, DMSO-d₆) δ ppm 7.62 (s, 1H), 7.57 (s, 1H), 7.47 (td, J=8.9, 3.0 Hz, 2H), 7.01 (ddd, J=11.5, 8.7, 2.8 Hz, 2H), 6.83-6.75 (m 2H), 5.39 (dd, J=9.7, 2.1 Hz, 1H), 4.48-4.37 (m, 4H), 4.10-3.98 (in, 214), 3.53-3.45 (m, 2H), 2.42 (ddd, J=14.1, 9.4, 3.0 Hz, 1H), 2.22 (d, J=12.6 Hz, 1H), 2.05-1.96 (m, 1H), 1.96-1.90 (m, 2H), 1.90-1.75 (m, 5H), 1.11 (t, J=7.0 Hz, 3H): MS (ESI⁺) m/z 616 (M+H)⁺.

Example 7: 4-({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)-4-oxobutanoic Acid (Compound 106)

A suspension of the product of Example 1L (14 mg, 0.026 mmol) and N,N-dimethylpyridin-4-amine (3.23 mg, 0.026 mmol) in N,N-dimethylformamide (0.5 mL) was cooled to 0° C. and treated with dihydrofuran-2,5-dione (5.29 mg, 0.053 mmol). The reaction mixture was stirred at 25° C. for 20 hours, then the temperature was raised to 40° C., and the reaction was stirred for 72 hours. The mixture was concentrated under reduced pressure, and the residue was purified on HPLC (Phenomenex® Luna® C18(2) 5 μm 100 Å AXIA™ column 250 mm×21.2 mm, flow rate 25 mL/minute, 10-80% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid in water)) to give the title compound (13 mg, 0.021 mmol, 78% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.21 (s, 1H), 7.60 (s, 1H), 7.52-7.41 (m, 3H), 7.00 (ddd, J=14.7, 11.4, 2.9 Hz, 2H), 6.79 (td d, =9.0, 2.8, 1.2 Hz, 2H), 5.31 (d, J=8.3 Hz, 114), 4.43 (d, J=2.9 Hz, 4H), 2.51-2.37 (m, 3H), 2.2.4 (t, J=10.3 Hz, 1H), 1.99-1.73 (in, 8H); MS (ESI⁺) m/z 630 (M+H)+.

Example 8: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Ethyl Carbonate (Compound 107)

To a suspension of the product of Example 1L (0.0500 g, 0.094 mmol) in pyridine (0.10 mL, 1.24 mmol) at room temperature was added ethyl carbonochloridate (10 μL, 0.105 mmol), and the mixture was stirred for 16 hours and then was heated to 50° C. for 8 hours. Another aliquot of ethyl carbonochloridate (10 μL, 0.105 mmol) was added. The mixture was stirred at 50° C. for 5 hours, and additional ethyl carbonochloridate (10 μL, 0.105 mmol) was again added. The mixture was stirred for 4 hours, diluted with ethyl acetate, washed with water and brine, dried (Na₂SO₄), and concentrated under reduced pressure. The residue was purified on silica gel (10% ethyl acetate/dichloromethane) to give the title compound (0.026 g, 0.044 mmol, 46% yield). ¹H NMR (400 MHz, DMSO-d₆) (5 ppm 7.66 (s, 1H), 7.63 (s, 1H), 7.46 (dt, J=9.9, 8.8 Hz, 2H), 6.99 (ddd, J=17.9, 11.4, 2.9 Hz, 2H), 6.84-6.75 (m, 2H), 5.26 (dd, J=9.2, 2.0 Hz, 1H), 4.51-4.37 (m, 4H), 4.16-3.99 (m, 2H), 2.40 (ddd, J=14.3, 9.3, 2.7 Hz, 1H), 2.34-2.22 (m, 1H), 2.00 (d, J=9.2 Hz, 1H), 1.97-1.70 (m, 7H), 1.21 (t, J=7.1 Hz, 3H); MS (ESI) m/z 598 (M−H)⁻.

Example 9: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Diethylcarbamate (Compound 108) Example 9A: N,N′-[(2S)-2-({[(2,5-dioxopyrrolidin-1-yl)oxy]carbonyl}oxy)bicyclo[2.2.2]octane-1,4-(4-chloro-3-fluorophenoxy)acetamide]

A mixture of the product of Example 1L (0.40 g, 0.756 mmol) and N,N′-disuccinimidyl carbonate (0.58 g, 2.27 mmol) in pyridine (1 mL) was heated to 50° C. for 4 hours. The reaction mixture was concentrated under reduced pressure. The concentrate was treated with brine and aqueous NaHCO₃, and extracted with ethyl acetate (2×). The combined organic layers were dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The residue was purified on a 40 g silica column using the Biotage Isolera™ One flash system eluted with heptanes/ethyl acetate (4:6 to 3:7) to provide the title compound (0.292 g, 58%). MS (EST⁺) m/z 670.2 (M+H)⁺.

Example 9B: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Diethylcarbamate

A solution of the product of Example 9A (0.061 g, 0.091 mmol) and diethylamine (38 μL, 0.367 mmol) in tetrahydrofuran (0.30 mL) was stirred at room temperature for 1 hour. The mixture was then diluted with ethyl acetate, washed with 1 N NaOH and brine, dried (Na₂SO₄) and concentrated under reduced pressure. The residue was purified on silica gel (20-25% ethyl acetate/dichloromethane) to give the title compound (0.024 g, 0.038 mmol, 42% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.69 (s, 1H), 7.60 (s, 1H), 7.46 (q, J=8.9 Hz, 2H), 7.00 (ddd, J=13.1, 11.4, 2.9 Hz, 2H), 6.79 (tdd, J=8.6, 2.9, 1.2 Hz, 2H), 5.15-5.08 (m, 1H), 4.42 (m, 4H), 3.19 (p, J=7.4 Hz, 4H), 2.36 (ddd, J=14.2, 9.2, 2.8 Hz, 1H), 2.17-1.69 (m, 9H), 1.04 (d, J=8.6 Hz, 6H); MS (ESI⁺) m/z 628.0 (M+H)⁺.

Example 10: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (phosphonooxy)acetate (Compound 109) Example 10A: benzyl[(di-tert-butoxyphosphoryl)oxy]acetate

To a solution of benzyl 2-hydroxyacetate (0.276 mL, 1.942 mmol) and a 0.45 M acetonitrile solution of 1H-tetrazole (12.5 mL, 5.63 mmol) was added di-tert-butyl diethylphosphoramidite (0.80 mL, 2.87 mmol), and the mixture was stirred at room temperature for 2 hours and then was cooled to 0° C. Hydrogen peroxide (1.0 mL, 9.79 mmol) was added, and the reaction was stirred for 90 minutes. The mixture was diluted with ethyl acetate (15 mL), washed with saturated Na₂SO₃ (10 mL) and brine (10 mL), dried (Na₂SO₄) and concentrated under reduced pressure. The residue was purified on silica gel (10% ethyl acetate/dichloromethane) to give the title compound (0.718 g, 2.0 mmol, quantitative). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.42-7.31 (m, 5H), 5.19 (s, 2H), 4.54 (d, J=9.7 Hz, 2H), 1.39 (s, 18H).

Example 10B: [(di-tert-butoxyphosphoryl)oxy]acetic Acid

A solution of the product of Example 10A (0.2376 g, 0.663 mmol) and 1 M sodium hydroxide (0.73 mL, 0.730 mmol) in methanol (0.75 mL) and tetrahydrofuran (0.75 mL) was stirred for 10 minutes, diluted with water (8 mL), and extracted with ethyl acetate (8 mL). The aqueous fraction was neutralized with 1 N HCl (0.8 mL), extracted with CH₂Cl₂ (3×6 mL), dried (Na₂SO₄), and concentrated to give the title compound (0.145 g, 0.54 mmol, 81% yield) as a clear oil. ¹H NMR (501 MHz, DMSO-d₆) (5 ppm 4.36 (d, J=9.0 Hz, 2H), 1.41 (d, J=0.5 Hz, 18H).

Example 10C: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl [(di-tert-butoxyphosphoryl)oxy]acetate

A solution the product of Example 10B (0.0277 g, 0.103 mmol), HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate) (0.0473 g, 0.124 mmol) and triethylamine (0.020 mL, 0.143 mmol) in N,N-dimethylformamide (0.30 mL) at room temperature was stirred for 15 minutes, and then the product of Example 1L (0.050 g, 0.095 mmol) was added. The mixture was stirred overnight, diluted with ethyl acetate, washed with water, 1 N NaOH, and brine, dried (Na₂SO₄), and concentrated under reduced pressure. The residue was purified on silica gel (25-50% ethyl acetate/dichloromethane) to give a ˜1:1 mixture of the product of Example 1L and the title compound (39.2 mg mixture, ˜31% yield product). This mixture was carried on without purification. LC/MS (APCI⁺) m/z 667 (M−2(t−Bu)+H)⁺.

Example 10D: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (phosphonooxy)acetate

A solution of the mixture from Example 10C (0.025 mmol) and 2,2,2-trifluoroacetic acid (40 μL, 0.523 mmol) in CH₂Cl₂ (0.20 mL) at room temperature was stirred for 1 hour. The mixture was concentrated and purified by preparative HPLC (Phenomenex® Luna® 250×30 mm 10 μm C18 column, 60 mL/minute, 15-95% gradient of acetonitrile in 0.1% aqueous trifluoroacetic acid) to give the product containing residual dimethyl sulfoxide. This material was diluted with ethyl acetate, washed with water, concentrated and dried under vacuum dried to give the title compound (0.014 g, 0.021 mmol, 41.7% yield). ¹H NMR (501 MHz, DMSO-d₆) ppm 7.63 (s, 1H), 7.58 (s, 1H), 7.46 (td, J=8.9, 6.3 Hz, 2H), 7.00 (ddd, J=14.0, 11.4, 2.9 Hz, 2H), 6.79 (ddd, J=12.8, 9.0, 2.9 Hz, 2H), 5.36 (d, J=9.1 Hz, 1H), 4.46-4.35 (m, 6H), 2.48-2.38 (m, 1H), 2.30-2.15 (m, 1H), 2.06-1.72 (m, 8H); MS (ESI) m/z 664.8 (M−H)⁻.

Example 11 (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (phosphonooxy)methyl Carbonate (Compound 110) Example 11A: O-(chloromethyl)S-propyl Carbonothioate

N,N-Diisopropylethylamine (33.1 mL, 189 mmol) and propane-1-thiol (16 mL, 172 mmol) were dissolved in methyl tert-butyl ether (345 mL) in a 1-L round-bottomed flask, and the flask was cooled to <5° C. before addition of chloromethyl carbonochloridate (16.08 mL, 181 mmol) dropwise via syringe. The reaction was stirred overnight while warming to room temperature. The resulting white suspension was treated with 100 mL of 1 M HCl (aqueous) and the layers were separated. The organic layer was washed with 1 M HCl (2×50 mL) and brine (50 mL) then dried over anhydrous sodium sulfate and concentrated in vacuo to give 29 g of the title compound as a colorless oil that was used without additional purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 5.77 (s, 2H), 2.90 (dd, J=7.6, 6.9 Hz, 2H), 1.71 (dt, J=14.5, 7.3 Hz, 2H), 1.01 (t, J=7.4 Hz, 3H).

Example 11B: O-(iodomethyl)S-propyl Carbonothioate

Example 11A (29.5 g, 175 mmol) was dissolved in acetone (200 mL) and sodium iodide (52.4 g, 350 mmol) was added. The reaction mixture was heated to 40° C. after wrapping the flask in foil. After 90 minutes, the flask was cooled to room temperature, and solid material was removed via filtration through a fritted funnel. Then the filtrate was concentrated under reduced pressure to approximately 30 mL. The solution was diluted with methyl tert-butyl ether (300 mL) and washed with water (3×50 mL) and saturated, aqueous sodium thiosulfate (2×25 mL), water (50 mL) and brine (50 mL), and then was dried over anhydrous sodium sulfate. The mixture was concentrated in vacuo to give a crude oil that was used without additional purification (44.8 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 5.99 (s, 2H), 2.95-2.85 (m, 2H), 1.70 (h, J=7.3 Hz, 2H), 1.01 (t, J=7.3 Hz, 3H).

Example 11C: Silver (1+) Dibenzyl Phosphate

Dibenzyl hydrogen phosphate (26.7 g, 96 mmol) was suspended in 300 mL of deionized water, and sodium hydroxide (1 M aqueous, 96 mL, 96 mmol) solution was added. The resulting suspension was stirred at room temperature for 30 minutes, at which point a complete dissolution resulted and a pH of 6-7 was achieved. A separate solution of silver(I) nitrate (17.12 g, 101 mmol) in 150 mL of water was added dropwise via addition funnel while stirring vigorously. The resulting white solid was isolated by filtration through a 600-mL fritted funnel. The isolated solid was washed with acetone (2×200 mL) and methyl tert-butyl ether (200 mL) and then was dried to constant weight in a vacuum oven at 70° C. to give 34.1 g of the title compound as a white solid, which was used without additional purification or characterization.

Example 11D: O-({[bis(benzyloxy)phosphoryl]oxy}methyl)S-propyl Carbonothioate

Example 11B (44.8 g, 172 mmol) was dissolved in acetonitrile (344 mL) and Example 11C (66.3 g, 172 mmol) was added in one portion. The resulting light yellow suspension was stirred at room temperature for 3 hours under N₂. Solid material was removed via filtration through a fritted funnel, and the filtrate was diluted with 100 mL of ethyl acetate and then refiltered. The filtrate was concentrated in vacuo to give the title compound as a light yellow oil that was used without additional purification (63.8 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 7.34 (d, J=2.3 Hz, 10H), 5.65 (d, J=13.9 Hz, 2H), 5.07 (d, J=7.9 Hz, 4H), 2.92-2.69 (m, 2H), 1.66 (hept, J=7.5 Hz, 2H), 0.97 (t, J=7.4 Hz, 3H); MS (ESI) m/z 411.1 (M+H)⁺.

Example 11E: {[bis(benzyloxy)phosphoryl]oxy}methyl Carbonochloridate

A 25-mL round-bottomed flask was charged with Example 11D (25.5 g, 62.1 mmol) and a stir bar. After cooling to <5° C. in an ice-water bath, sulfuryl chloride (6.06 mL, 74.6 mmol) was added dropwise. The solution was stirred for 30 minutes at the same temperature and then was allowed to warm to room temperature for 1 hour, at which point ¹H NMR analysis showed complete conversion. Volatile material was removed in vacuo at 35° C. The resulting crude oil was loaded onto a 330 g silica gel column and purified via flash chromatography, eluted with 0:100 to 50:50 ethyl acetate:heptanes over 15 minutes and then isocratically 50:50 ethyl acetate:heptanes for 5 minutes to give 12.5 g of the title compound as a light yellow oil. ¹H NMR (500 MHz, CDCl₃) (5 ppm 7.41-7.30 (m, 10H), 5.62 (d, J=14.5 Hz, 2H), 5.08 (d, J=8.4 Hz, 4H).

Example 11F: {[bis(benzyloxy)phosphoryl]oxy}methyl (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Carbonate

A suspension of Example 11E (2.0670 g, 5.58 mmol) and N,N-dimethylpyridin-4-amine (0.69 g, 5.67 mmol) in CH₂Cl₂ (15 mL) was stirred for 2 minutes, and the product of Example 1L (1.5 g, 2.83 mmol) and then triethylamine (1.0 mL, 7.17 mmol) were added. The mixture was stirred for 1 hour and then was quenched with water and concentrated under reduced pressure. The residue was diluted with ethyl acetate and then washed with 1 N HCl (aqueous), water, and brine. The organic fraction was dried (Na₂SO₄) and concentrated under reduced pressure. The residue was purified on silica gel (50-90% ethyl acetate/heptane, 40 g Teledyne Isco RediSep Rf Gold®) to give a white solid, which was crystallized from ethyl acetate (˜0.5 volume) and tert-butyl methyl ether (˜4 volumes) to give the title compound (1.42 g, 1.64 mmol, 58% yield). ¹H NMR (501 MHz, DMSO-d₆) δ ppm 7.68 (s, 1H), 7.65 (s, 1H), 7.49-7.44 (m, 1H), 7.42 (d, J=8.9 Hz, 1H), 7.39-7.30 (m, 10H), 7.02 (dd, J=11.4, 2.8 Hz, 1H), 6.94 (dd, J=11.4, 2.9 Hz, 1H), 6.80 (ddd, J=8.9, 2.9, 1.1 Hz, 1H), 6.74 (ddd, J=9.0, 2.9, 1.1 Hz, 1H), 5.65 (dd, J=14.3, 5.7 Hz, 1H), 5.58 (dd, J=12.9, 5.7 Hz, 1H), 5.39 (dd, J=9.4, 2.2 Hz, 1H), 5.05 (dd, J=8.2, 1.8 Hz, 4H), 4.50-4.33 (m, 4H), 2.49-2.43 (m, 1H), 2.29 (m, 8.1 Hz, 1H), 2.03-1.92 (m, 2H), 1.92-1.73 (m, 6H); LC/MS (APCI⁺) m/z 863 (M+H)⁺.

Example 11G: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (phosphonooxy)methyl Carbonate

To a solution of Example 11F (0.030 g, 0.035 mmol) in tetrahydrofuran (1.0 mL) and 4 N HCl in dioxane (0.019 mL, 0.077 mmol) was added to 5% Pd/C (wet JM #9) (5.91 mg, 0.025 mmol) in a 20 mL Barnstead Hast C reactor. The reactor was purged with argon, and the mixture was stirred at 1200 RPM under 50 psi of hydrogen at 25° C. The mixture was vented after 1 hour and filtered. The filtrate was concentrated under reduced pressure and azeotroped with toluene. The resulting material was slurried with 1:1 methyl tert-butyl ether:heptane and solids were isolated via filtration. The solids were dissolved in acetone and concentrated to remove previous solvents. The material was then dissolved in acetonitrile (5 mL) and water (1 mL) and lyophilized to give the title compound (1.02 g, 1.49 mmol, 92% yield). ¹H NMR (400 MHz, DMSO-d₆) (5 ppm 7.66 (d, J=6.6 Hz, 2H), 7.47 (td, J=8.9, 5.9 Hz, 2H), 7.01 (ddd, J=15.7, 11.4, 2.9 Hz, 2H), 6.79 (td, J=9.5, 2.8 Hz, 2H), 5.47 (ddd, J=24.8, 13.2, 5.5 Hz, 2H), 5.35 (dd, J=9.3, 2.2 Hz, 1H), 4.51-4.37 (m, 4H), 2.49-2.40 (m, 1H), 2.32 (m, 1H), 2.06-1.69 (m, 8H); MS (ESI⁻) m/z 680.6 (M−H)⁻.

Example 12: [4-({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)-4-oxobutyl]phosphonic Acid (Compound 111)

To a suspension of 4-phosphonobutanoic acid (0.0509 g, 0.303 mmol) in CH₂Cl₂ (0.60 mL) and catalytic N,N-dimethylformamide (1 drop) at room temperature was added oxalyl chloride (0.030 mL, 0.344 mmol), and the mixture was stirred for 1 hour. The mixture was then concentrated under reduced pressure, and the residue was dissolved in N,N-dimethylformamide (0.60 mL). The product of Example 1L (0.158 g, 0.30 mmol) was added, followed by N,N-dimethylpyridin-4-amine (0.037 g, 0.30 mmol) and triethylamine (0.060 mL, 0.430 mmol). The reaction was stirred for 2 hours and then was warmed to 70° C. and allowed to stir for 4 days. The mixture was partitioned between ethyl acetate and 1 N HCl (1 mL, aqueous). The organic fraction was washed with water and brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by preparative LC (YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, 140 mL/minute, 3-100% gradient of acetonitrile in 25 mM ammonium bicarbonate buffer, adjusted to pH 10 with concentrated aqueous NH₄OH) to give the title compound (0.025 g, 0.037 mmol, 12% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.95 (s, 1H), 7.65 (s, 1H), 7.44 (dt, J=13.6, 8.9 Hz, 2H), 6.99 (ddd, J=14.6, 11.4, 2.9 Hz, 2H), 6.79 (ddd, J=9.1, 6.1, 2.8 Hz, 2H), 5.32 (dd, J=9.3, 2.2 Hz, 1H), 4.46 (d, J=2.6 Hz, 2H), 4.43 (s, 2H), 2.48-2.18 (m, 5H), 2.05-1.64 (m, 9H), 1.44 (m, 2H); MS (ESI) m/z 677.0 (M−H)⁻.

Example 13: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (benzyloxy)acetate (Compound 112)

To a suspension of the product of Example 1L (0.3 g, 0.567 mmol), 2-(benzyloxy)acetic acid (0.141 g, 0.850 mmol) and N,N-diisopropylethylamine (0.30 mL, 1.70 mmol) in N,N-dimethylformamide (5 mL) was added 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (0.648 g, 1.71 mmol), and the mixture was stirred at ambient temperature for 16 hours. Water was added, and the aqueous layer was extracted with dichloromethane (2×100 mL). The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by HPLC (Phenomenex® Luna® C18(2) 10 μm 100 Å AXIA™ column (250 mm×50 mm). A 30-100% gradient of acetonitrile (A) and 0.1% trifluoroacetic acid in water (B) is used over 25 minutes, at a flow rate of 50 mL/minute) to give 380 mg of title compound. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.66 (d, J=19.7 Hz, 1H), 7.52-7.33 (m, 2H), 7.37-7.25 (m, 4H), 6.99 (ddd, J=17.3, 11.4, 2.9 Hz, 2H), 6.79 (dddd, J=18.7, 8.9, 2.8, 1.2 Hz, 2H), 5.46 (dd, J=9.6, 2.1 Hz, 1H), 4.59-4.50 (m, 2H), 4.52-4.34 (m, 4H), 4.14 (d, J=1.4 Hz, 2H), 2.44 (td, J=10.6, 9.1, 5.4 Hz, 1H), 2.26 (d, J=12.3 Hz, 1H), 2.02 (t, J=9.2 Hz, 1H), 1.98-1.84 (m, 4H), 1.84 (d, J=11.9 Hz, 3H); MS (ESI+) m/z 694.0 (M+H)⁺.

Example 14: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl [3-(phosphonooxy)phenyl]acetate (Compound 113) Example 14A: methyl {3-[(di-tert-butoxyphosphoryl)oxy]phenyl}acetate

To a solution of methyl 2-(3-hydroxyphenyl)acetate (2.5 g, 15 mmol) and 1H-tetrazole (67 mL, 30 mmol, 0.45 M in acetonitrile) in anhydrous CH₂Cl₂ (20 mL) at 0° C. was added di-tert-butyl diisopropylphosphoramidite (7.1 mL, 23 mmol). The resulting solution was allowed to warm to ambient temperature and was stirred for 4.5 hours. The reaction mixture was cooled to 0° C., and t-butyl hydroperoxide (70% aqueous solution, 6.2 mL, 45 mmol) was added. The resulting mixture was allowed to warm to ambient temperature, was stirred for 1 hour, and then was washed with saturated Na₂S₂O₃ solution (50 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (SiO₂, 0-50% ethyl acetate/heptanes) to afford the title compound as a colorless oil (4.77 g, 13.3 mmol, 89% yield). ¹H NMR (400 MHz, DMSO-d₆) ppm 7.32 (t, J=7.8 Hz, 1H), 7.15-7.04 (m, 3H), 3.70 (s, 2H), 3.61 (s, 3H), 1.44 (d, J=0.7 Hz, 18H).

Example 14B: {3-[(di-tert-butoxyphosphoryl)oxy]phenyl}acetic Acid

To a cooled (0° C. ice bath) solution of the product of Example 14A (4.77 g, 13.3 mmol) in tetrahydrofuran (15 mL) and water (5 mL) was added lithium hydroxide (0.96 g, 40 mmol). The resulting mixture was stirred at ambient temperature for 2 hours and then 1.0 M HCl was added to acidify the reaction mixture to pH 3.0. The reaction mixture was extracted with ethyl acetate (50 mL). The organic fraction was washed with water (50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to yield the titled intermediate as a yellow oil (3 g, 8.7 mmol, 65% yield), which was carried forward without purification. ¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.32 (s, 1H), 7.34-7.28 (m, 1H), 7.11 (dq, J=4.0, 2.6, 1.9 Hz, 1H), 7.09-7.04 (m, 2H), 3.57 (s, 2H), 1.46-1.43 (m, 18H).

Example 14C: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl {3-[(di-tert-butoxyphosphoryl)oxy]phenyl}acetate

To a mixture of the product of Example 1L (1 g, 2 mmol) and the product of Example 14B (0.68 g, 2.0 mmol) in N,N-dimethylformamide (10.73 mL) was added triethylamine (1.1 mL, 7.6 mmol) followed by 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (HATU, 0.79 g, 2.1 mmol). This mixture was allowed to stir at ambient temperature 20 hours, and then the reaction mixture was filtered, and concentrated under reduced pressure. The residue was directly purified by preparative HPLC [Waters XBridge™ C18 5 μm OBD column, 30×100 mm, flow rate 40 mL/minute, 5-100% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid)] to provide the title intermediate (0.49 g, 0.58 mmol, 31% yield). ¹⁻H NMR (501 MHz, DMSO-d₆) δ ppm 7.56 (s, 1H), 7.49 (s, 1H), 7.43 (dt, J=18.7, 8.9 Hz, 2H), 7.27 (t, J=7.9 Hz, 1H), 7.10-7.02 (m, 3H), 6.96 (ddd, J 26.8, 11.4, 2.9 Hz, 2H), 6.77 (ddd, J=8.8, 2.8, 1.1 Hz, 1H), 6.71 (ddd, J=9.0, 2.9, 1.1 Hz, 1H), 5.32 (dd, J=9.6, 2.3 Hz, 1H), 4.40 (s, 2H), 4.37 (d, J=15.0 Hz, 1H), 4.29 (d, J=14.6 Hz, 1H), 3.64 (d, J=3.0 Hz, 2H), 2.40 (ddd, J=13.9, 9.0, 2.6 Hz, 1H), 2.25-2.16 (m, 1H), 1.89-1.69 (m, 8H), 1.40 (d, J=2.5 Hz, 18H).

Example 14D: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl[3-(phosphonooxy)phenyl]acetate

To a solution of the product of Example 14C (0.49 g, 0.58 mmol) in CH₂Cl₂ (9.6 mL) was added trifluoroacetic acid (2 mL). The reaction mixture stirred for 1 hour and then was concentrated under reduced pressure. The resulting solids were dried under vacuum for 2 days to afford the title compound (0.43 g, 0.58 mmol, quantitative yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.61 (d, J=8.0 Hz, 2H), 7.45 (dt, J=15.4, 8.9 Hz, 2H), 7.26 (t, J=7.9 Hz, 1H), 7.14 (d, J=1.9 Hz, 1H), 7.08-7.02 (m, 3H), 7.02-6.91 (m, 2H), 6.76 (ddt, J=35.0, 9.1, 1.8 Hz, 3H), 5.38 (dd, J=9.5, 2.3 Hz, 1H), 4.43 (s, 2H), 4.40 (d, J=14.5 Hz, 1H), 4.24 (d, J=14.5 Hz, 1H), 3.65 (s, 2H), 2.41 (dd, J=14.1, 9.5 Hz, 1H), 2.29 (td, J=11.9, 5.7 Hz, 1H), 1.96-1.72 (m, 8H); MS (EST⁺) m/z 744 (M+H)⁺.

Example 15: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 3-[2,4-dimethyl-6-(phosphonooxy)phenyl]-3-methylbutanoate (Compound 114) Example 15A: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 3-(2-{[bis(benzyloxy)phosphoryl]oxy}-4,6-dimethylphenyl)-3-methylbutanoate

To a solution of the product of Example 1L (0.150 g, 0.283 mmol), 4-dimethylaminopyridine (0.042 g, 0.340 mmol), and 3-(2-((bis(benzyloxy)phosphoryl)oxy)-4,6-dimethylphenyl)-3-methylbutanoic acid (0.178 g, 0.368 mmol, CAS #153910-62-4, Bioorganic & Medicinal Chemistry Letters, 1993, 3(8), 1761-1766) in N,N-dimethylformamide (4 mL) was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.087 g, 0.45 mmol), and the mixture was stirred overnight. The reaction mixture was then quenched with brine and saturated, aqueous NaHCO₃, and extracted with ethyl acetate (2×). The combined organic layers were washed with brine, dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The residue was purified on a 40 g silica column using a Biotage Isolera™ One flash system eluted with heptanes/ethyl acetate (4:6 to 3:7) to provide 0.101 g of the title compound (36%). MS (ESI⁺) m/z 993.4 (M+H)⁺.

Example 15B: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 3-[2,4-dimethyl-6-(phosphonooxy)phenyl]-3-methylbutanoate

A mixture of Example 15A (97.0 mg, 0.098 mmol) and trifluoroacetic acid (0.15 mL, 1.95 mmol) in CH₂Cl₂ (0.4 mL) was stirred for 3 days. The reaction mixture was concentrated under reduced pressure and purified by reverse-phase HPLC performed on a Phenomenex® Luna® C18 column (250×30 mm, 10 μm particle size) using a gradient of 20% to 100% acetonitrile:0.1% aqueous trifluoroacetic acid over 26 minutes at a flow rate of 50 mL/minute to provide the title compound (57.5 mg, 58%). ¹H NMR (400 MHz, CD₃OD) δ ppm 7.35 (dt, J=19.5, 8.7 Hz, 2H), 7.08 (s, 1H), 6.91 (dd, J=11.0, 2.8 Hz, 1H), 6.87-6.77 (m, 2H), 6.72 (ddd, J=8.9, 2.8, 1.2 Hz, 1H), 6.63-6.53 (m, 1H), 5.33 (dd, J=9.5, 1.9 Hz, 1H), 4.41 (s, 2H), 4.32 (a of ab, J=14.8 Hz, 1H), 4.14 (b of ab, J=14.8 Hz, 1H), 3.04 (s, 2H), 2.49-2.38 (m, 4H), 2.26 (td, J=11.5, 11.0, 5.8 Hz, 1H), 2.05-1.69 (m, 8H), 1.63 (s, 3H), 1.58 (s, 3H); MS (ESI⁺) m/z 813.1 (M+H)⁺.

Example 16: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl [2-(phosphonooxy)phenyl]acetate (Compound 115) Example 16A: methyl (2-{[bis(benzyloxy)phosphoryl]oxy}phenyl)acetate

Dibenzyl diisopropylphosphoramidite (1.35 mL, 3.61 mmol) was added to a stirred solution of methyl 2-(2-hydroxyphenyl)acetate (0.48 g, 2.89 mmol; Advanced ChemBlocks) and 1H-tetrazole (0.45 M in acetonitrile, 16.05 mL) in dimethylacetamide (5.8 mL). The reaction mixture was stirred at ambient temperature for 1 hour and was then cooled to 0° C. Hydrogen peroxide solution (30 weight %, 0.738 mL) was added dropwise. The reaction mixture was slowly warmed up to ambient temperature over a period of 30 minutes and stirred for another 2 hours before being cooled back to 0° C. Saturated, aqueous sodium bicarbonate solution (30 mL) was added in one portion, and the resulting mixture was partitioned between ethyl acetate (2×50 mL) and aqueous sodium thiosulfate solution (1.0 M, 30 mL). The organic phases were combined, dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by preparative HPLC [YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, flow rate 140 mL/minute, 5-100% gradient of acetonitrile in buffer (0.025 M aqueous ammonium bicarbonate, adjusted to pH 10 with ammonium hydroxide)] to give the title compound (1.12 g, 2.63 mmol, 91% yield). MS (ESI⁺) m/z 427 (M+H)⁺.

Example 16B: (2-{[bis(benzyloxy)phosphoryl]oxy}phenyl)acetic Acid

The product of Example 16A (1.12 g, 2.63 mmol) was stirred in a solvent mixture of water (10.4 mL) and tetrahydrofuran (26.1 mL) at 0° C. Aqueous LiOH solution (0.3 M, 26.1 mL) was added dropwise over a period of 2 minutes. The reaction mixture was stirred at 0° C. for 1 hour and was slowly allowed to warm to ambient temperature over a period of 30 minutes. Aqueous HCl solution (1.0 M) was added to adjust the pH to 3.0. The resulting mixture was extracted with ethyl acetate. The organic layer was first washed with water and then with brine, dried over sodium sulfate, and concentrated under reduced pressure to give the title compound (1.15 g, 2.79 mmol, quantitative yield). MS (ESI⁺) m/z 413 (M+H)⁺.

Example 16C: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (2-{[bis(benzyloxy)phosphoryl]oxy}phenyl)acetate

Bis(tetramethylene)fluoroformamidinium hexafluorophosphate (BTFFH, 564 mg, 1.78 mmol) and the product of Example 16B (490 mg, 1.19 mmol) were charged to a 20 mL vial. A solvent mixture of dichloromethane (4.75 mL) and N,N-diisopropylethylamine, (0.93 mL) was added in one portion. The resulting mixture was stirred at ambient temperature for 30 minutes. The product of Example 1L (755 mg, 1.43 mmol) and N,N-dimethylpyridin-4-amine (36.3 mg, 0.30 mmol) were added sequentially, and the vial was sealed and stirred at ambient temperature for 18 hours. The resulting reaction mixture was partitioned between water (10 mL) and dichloromethane (10 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC [YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, flow rate 140 mL/minute, 5-100% gradient of acetonitrile in buffer (0.025 M aqueous ammonium bicarbonate, adjusted to pH 10 with ammonium hydroxide)] to give the title compound (0.22 g, 0.24 mmol, 20% yield). MS (APCI⁺) m/z 923 (M+H)⁺.

Example 16D: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl [2-(phosphonooxy)phenyl]acetate

Palladium on carbon (43 mg, 0.19 mmol; 5 weight % loading (dry basis)) was added to a solution of the product of Example 16C (212 mg, 0.23 mmol) in tetrahydrofuran (9 mL) in a 20 mL Barnstead Hast C reactor followed by addition of HCl solution (4.0 M in dioxane, 0.126 mL). The reaction mixture was stirred for 8.5 minutes at 25° C. under hydrogen (50 psi). The resulting mixture was then filtered and concentrated under reduced pressure. The residue was taken up in methanol (3 mL) and purified by preparative HPLC [YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, flow rate 140 mL/minute, 0-100% gradient of acetonitrile in buffer (0.025 M aqueous ammonium bicarbonate)] to give the title compound (105 mg, 0.14 mmol, 56% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.35 (s, 1H), 7.61 (s, 1H), 7.48 (t, J=8.9 Hz, 1H), 7.36 (d, J=8.1 Hz, 1H), 7.20-7.13 (m, 2H), 7.10 (td, J=7.8, 1.8 Hz, 1H), 7.03 (dd, J=11.4, 2.8 Hz, 1H), 6.97 (dd, J=11.6, 2.9 Hz, 1H), 6.89 (t, J=7.4 Hz, 1H), 6.85-6.77 (m, 1H), 6.74-6.66 (m, 1H), 5.37 (d, J=9.0 Hz, 1H), 4.57-4.46 (m, 2H), 4.44 (s, 2H), 3.83-3.45 (m, 2H), 2.49-2.42 (m, 1H), 2.38-2.29 (m, 1H), 2.08-1.58 (m, 8H); MS (ESI⁺) m/z 743 (M+H)⁺.

Example 17: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl methyl{3-[(phosphonooxy)methyl]pyridin-2-yl}carbamate (Compound 116) Example 17A: prop-2-en-1-yl[3-(hydroxymethyl)pyridin-2-yl]methylcarbamate

Allyl chloroformate (2.90 mL, 27.1 mmol) was added to a suspension of (2-(methylamino)pyridin-3-yl)methanol (3.0 g, 21.7 mmol) in a mixture of ethyl acetate (15 mL) and saturated, aqueous sodium bicarbonate (15 mL), and the resulting reaction mixture was stirred at ambient temperature for 16 hours. The mixture was transferred to a separatory funnel, and the layers were separated. The organic layer was washed with water and brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the title compound (4.0 g, 18 mmol, 83%). MS (APCI⁺) m/z 223 (M+H)⁺.

Example 17B: prop-2-en-1-yl (3-{[(di-tert-butoxyphosphoryl)oxy]methyl}pyridin-2-yl)methylcarbamate

The reaction and purification conditions described in Example 16A, substituting di-tert-butyl diisopropylphosphoramidite for dibenzyl diisopropylphosphoramidite, and the product of Example 17A for methyl 2-(2-hydroxyphenyl)acetate gave the title compound. MS (APCI⁺) m/z 415 (M+H)⁺.

Example 17C: di-tert-butyl[2-(methylamino)pyridin-3-yl]methyl Phosphate

The product of Example 17B (750 mg, 1.81 mmol) was dissolved in a solvent mixture of dichloromethane (5 mL) and ethyl acetate (5 mL). 1,3-Dimethylbarbituric acid (367 mg, 2.35 mmol) and tetrakis(triphenylphosphine)palladium(O) (42 mg, 0.036 mmol) were added sequentially. The resulting reaction mixture was stirred at ambient temperature for 1 hour, concentrated, and then directly purified by flash chromatography (silica gel, 20-100% ethyl acetate in heptane) to give the title compound (0.3 g, 0.91 mmol, 50% yield). MS (APCI⁺) m/z 331 (M+H)⁺.

Example 17D: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl carbonochloridate

Pyridine (0.087 mL, 1.07 mmol) was added to a stirred mixture the product of Example 1L (515 mg, 0.97 mmol) in dichloromethane (4 mL) at ambient temperature. A solution of triphosgene (101 mg, 0.34 mmol) in dichloromethane (0.5 mL) was added dropwise over a period of 3 minutes. The resulting mixture was stirred at ambient temperature for 18 hours and then was directly purified via flash chromatography (SiO₂, 20% to 80% acetone in heptane) to give the title compound (0.4 g, 0.68 mmol, 70% yield). ¹H NMR (501 MHz, DMSO-d₆) δ ppm 7.50 (s, 1H), 7.46 (t, J=8.9 Hz, 1H), 7.45 (t, J=8.9 Hz, 1H), 7.28 (s, 1H), 7.03 (dd, J=11.5, 2.9 Hz, 1H), 7.00 (dd, J=11.4, 2.8 Hz, 1H), 6.83-6.75 (m, 2H), 4.44 (s, 2H), 4.41 (s, 2H), 4.01 (dd, J=9.6, 3.2 Hz, 1H), 2.24 (ddd, J=12.5, 9.4, 2.5 Hz, 1H), 2.09-2.02 (m, 1H), 1.98-1.68 (m, 8H).

Example 17E: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl methyl{3-[(phosphonooxy)methyl]pyridin-2-yl}carbamate

The product of Example 17C (27.9 mg, 0.084 mmol) was dissolved in tetrahydrofuran (5 mL). N,N-Diisopropylethylamine (0.024 mL, 0.135 mmol) was added followed by the product of Example 17D (50 mg, 0.084 mmol). The reaction mixture was stirred at ambient temperature for 18 hours and was then concentrated under reduced pressure. The resulting residue was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (2.0 mL, 26 mmol) was added in one portion. After stirring at ambient temperature for 30 minutes, the reaction mixture was concentrated under reduced pressure and then purified by preparative HPLC [YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, flow rate 140 mL/minute, 5-100% gradient of acetonitrile in buffer (0.025 M aqueous ammonium bicarbonate, adjusted to pH 10 with ammonium hydroxide)] to give the title compound (28 mg, 0.036 mmol, 43% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.31 (dd, J=4.8, 1.9 Hz, 1H), 8.02-7.97 (m, 1H), 7.42-7.27 (m, 3H), 7.02-6.90 (m, 2H), 6.84-6.75 (m, 2H), 5.37-4.94 (m, 2H), 4.70-4.59 (m, 2H), 4.58-4.49 (m, 1H), 4.36 (s, 2H), 3.13 (s, 3H), 2.37-2.26 (m, 2H), 1.94-1.16 (m, 8H); MS (EST⁺) m/z 773 (M+H)⁺.

Example 18: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl [2-(phosphonooxy)ethyl]carbamate (Compound 117) Example 18A: tert-butyl (2-{[bis(benzyloxy)phosphoryl]oxy}ethyl)carbamate

To a solution of tert-butyl (2-hydroxyethyl)carbamate (2.4 mL, 16 mmol) and 1H-tetrazole (69 mL, 31 mmol, 0.45 M in acetonitrile) in anhydrous CH₂Cl₂ (31 mL) at 0° C. was added dibenzyl diisopropylphosphoramidite (7.8 mL, 23 mmol), and the resulting solution was allowed to warm to ambient temperature and stirred overnight. The solution was then cooled to 0° C., and t-butyl hydroperoxide (70% aqueous solution, 6.4 mL, 47 mmol) was added. The resulting mixture was allowed to warm to ambient temperature, stirred for 1 hour and then was washed with saturated aqueous Na₂S₂O₃ solution (50 mL), followed by water (50 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the title intermediate (5 g, 12 mmol, 76% yield), which was used without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.43-7.33 (m, 10H), 5.06 (dd, J=13.4, 8.6 Hz, 4H), 4.02-3.88 (m, 2H), 3.21 (q, J=5.9 Hz, 2H), 1.37 (d, J=3.8 Hz, 9H); MS (ESI⁺) m/z 322 (M−Boc+H)⁺.

Example 18B: 2-aminoethyl Dibenzyl Phosphate

To a cooled (0° C. ice bath) solution of the product of Example 18A (0.1 g, 0.24 mmol) in CH₂Cl₂ (4 mL) was added trifluoroacetic acid (0.8 mL). The ice bath was removed and the reaction mixture stirred for 1 hour at ambient temperature. The solution was then concentrated under reduced pressure to afford the deprotected phosphate intermediate (0.058 g, 0.18 mmol, 75% yield), which was used in next step without purification. MS (ESI⁺) m/z 322 (M+H)⁺.

Example 18C: (28)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (2-{[bis(benzyloxy)phosphoryl]oxy}ethyl)carbamate

To a solution of the product of Example 9A (0.08 g, 0.12 mmol) and the product of Example 18B (0.15 g, 0.48 mmol) in tetrahydrofuran (0.4 mL) was added triethylamine (0.03 mL, 0.24 mmol). This mixture stirred at ambient temperature for 16 hours. Additional triethylamine (0.1 mL) was added, and the reaction mixture stirred for another 3 hours. Then the reaction mixture was concentrated under reduced pressure, and the residue was diluted with N,N-dimethylformamide (1 mL). The material was filtered and purified by preparative HPLC [Waters XBridge™ C18 5 μm OBD column, 50×100 mm, flow rate 90 mL/minute, 5-100% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid)] to afford the title intermediate (0.057 g, 0.064 mmol, 54% yield). ¹H NMR (501 MHz, DMSO-d₆) δ ppm 7.60 (d, J=14.2 Hz, 2H), 7.51-7.36 (m, 4H), 7.35 (q, J=4.4, 3.1 Hz, 8H), 7.30-7.18 (m, 1H), 7.01 (ddd, J=11.9, 9.1, 2.9 Hz, 2H), 6.84-6.74 (m, 2H), 5.12 (d, J=9.4 Hz, 1H), 5.01 (dd, J=8.0, 2.7 Hz, 4H), 4.41 (d, J=7.2 Hz, 3H), 4.36 (d, J=14.5 Hz, 2H), 3.97 (q, J=6.4 Hz, 2H), 3.27-3.16 (m, 2H), 2.32 (d, J=11.6 Hz, 1H), 2.05 (d, J=21.4 Hz, 4H), 1.83 (dt, J=43.6, 12.6 Hz, 4H); MS (EST⁺) m/z 876 (M+H)⁺.

Example 18D: (28)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl [2-(phosphonooxy)ethyl]carbamate

A mixture of the product of Example 18C (0.117 g, 0.133 mmol) and trifluoroacetic acid (0.205 mL, 2.66 mmol) in CH₂Cl₂ (0.55 mL) was stirred overnight. The reaction mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC [Waters XBridge™ C18 5 μm OBD column, 50×100 mm, flow rate 90 mL/minute, 5-100% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid)] to afford the title compound as a colorless oil (0.056 g, 0.080 mmol, 60% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.62 (d, J=15.0 Hz, 2H), 7.47 (td, J=8.8, 4.5 Hz, 2H), 7.26 (t, J=5.7 Hz, 1H), 7.02 (dt, J=11.3, 3.5 Hz, 2H), 6.81 (dd, J=8.9, 2.9 Hz, 2H), 5.11 (d, J=9.2 Hz, 1H), 4.42 (d, J=9.5 Hz, 4H), 3.82 (d, J=6.7 Hz, 2H), 3.20 (dq, J=12.8, 6.8 Hz, 2H), 2.38-2.28 (m, 1H), 2.14-1.96 (m, 4H), 1.84 (dd, J=37.8, 12.9 Hz, 5H); MS (ESI⁺) m/z 696 (M+H)⁺.

Example 19: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl [3-(phosphonooxy)propyl]carbamate (Compound 118) Example 19A: prop-2-en-1-yl (3-hydroxypropyl)carbamate

Allyl chloroformate (1.19 mL, 11.2 mmol) was added to a suspension of 3-aminopropan-1-ol (0.67 g, 8.92 mmol) in a solvent mixture of ethyl acetate (10 mL) and saturated aqueous sodium bicarbonate (10 mL). The reaction was stirred at ambient temperature for 3 hours. Water (30 mL) was added, and the layers were separated. The aqueous layer was washed again with a mixture of chloroform (30 mL) and 2-propanol (10 mL). All the organic layers were combined and dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title compound (1.15 g, 7.22 mmol, 81% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.12 (t, J=5.6 Hz, 1H), 5.90 (ddt, J=17.2, 10.6, 5.3 Hz, 1H), 5.26 (dq, J=17.3, 1.8 Hz, 1H), 5.16 (dq, J=10.6, 1.5 Hz, 1H), 4.49-4.42 (m, 2H), 3.40 (t, J=6.3 Hz, 2H), 3.03 (q, J=6.6 Hz, 2H), 1.55 (p, J=6.6 Hz, 2H); MS (APCI⁺) m/z 160 (M+H)⁺.

Example 19B: prop-2-en-1-yl (3-{[bis(benzyloxy)phosphoryl]oxy}propyl)carbamate

The reaction and purification conditions described in Example 16A, substituting the product of Example 19A for methyl 2-(2-hydroxyphenyl)acetate, gave the title compound. MS (EST⁺) m/z 420 (M+H)⁺.

Example 19C: 3-aminopropyl Dibenzyl Phosphate

The product of Example 19B (0.37 g, 0.88 mmol) was stirred in a solvent mixture of dichloromethane (2.0 mL) and ethyl acetate (2.0 mL) at ambient temperature. 1,3-Dimethylbarbituric acid (0.18 g, 1.14 mmol) was added followed by tetrakis(triphenylphosphine)palladium(O) (20 mg, 0.018 mmol). The reaction mixture was stirred at ambient temperature for 1 hour and was then concentrated under reduced pressure. The residue was taken up in N,N-dimethylformamide (3 mL), filtered through a glass microfiber frit and purified by preparative HPLC [YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, flow rate 140 mL/minute, 5-100% gradient of acetonitrile in buffer (0.025 M aqueous ammonium bicarbonate, adjusted to pH 10 with ammonium hydroxide)] to give the title compound (0.26 g, 0.78 mmol, 89% yield). MS (EST⁺) m/z 336 (M+H)⁺.

Example 19D: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (3-{[bis(benzyloxy)phosphoryl]oxy}propyl)carbamate

To a stirred suspension of the product of Example 19C (167 mg, 0.50 mmol) in ethyl acetate (15 mL) and saturated, aqueous sodium bicarbonate (15 mL) was added the product of Example 17D (200 mg, 0.34 mmol) in one portion. The resulting mixture was stirred at ambient temperature for 1 hour, and then the layers were separated. The organic layer was washed with water and brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title compound (0.27 g, 0.30 mmol, 90% yield). MS (ESI⁺) m/z 890 (M+H)⁺.

Example 19E: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl [3-(phosphonooxy)propyl]carbamate

Trifluoroacetic acid (2.0 mL, 26.0 mmol) was combined with the product of Example 19D (260 mg, 0.29 mmol), and the mixture was stirred at 50° C. for 1 hour. The reaction mixture was cooled to ambient temperature and then was concentrated under reduced pressure. The residue was taken up in methanol (5 mL), filtered through a glass microfiber frit and purified by preparative HPLC [custom packed YMC TriArt™ C18 Hybrid 20 μm plastic column, 25×70 mm, flow rate 70 mL/minute, 5-100% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid)] to give the title compound (155 mg, 0.22 mmol, 75% yield). ¹H NMR (400 MHz, DMSO-d6) (5 ppm 7.66 (s, 1H), 7.60 (s, 1H), 7.51-7.41 (m, 2H), 7.21 (t, J=5.7 Hz, 1H), 7.08-6.94 (m, 2H), 6.86-6.74 (m, 2H), 5.17-5.04 (m, 1H), 4.50-4.34 (m, 4H), 3.83 (q, J=6.7 Hz, 2H), 3.13-2.96 (m, J=6.9 Hz, 2H), 2.40-2.27 (m, 1H), 2.19-1.61 (m, 11H); MS (APCI⁺) m/z 710 (M+H)⁺.

Example 20: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 3-(phosphonooxy)propyl Carbonate (Compound 119) Example 20A: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 3-bromopropyl Carbonate

A mixture of Example 9A (0.120 g, 0.179 mmol), 4-dimethylaminopyridine (0.022 g, 0.179 mmol), 3-bromo-1-propanol (0.033 mL, 0.358 mmol), and triethylamine (0.027 mL, 0.197 mmol) in tetrahydrofuran (2 mL) was stirred overnight. The reaction mixture was quenched with brine and extracted with ethyl acetate (2×). The combined organic layers were dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The residue was purified on a 12 g silica column using the Biotage Isolera™ One flash system eluted with dichloromethane/ethyl acetate (9:1 to 7:3) to provide the title compound (61.0 mg, 49%). MS (ESI⁺) m/z 695.1 (M+H)⁺.

Example 20B: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 3-[(di-tert-butoxyphosphoryl)oxy]propyl Carbonate

A mixture of Example 20A (44.5 mg, 0.064 mmol) and tetra-N-butylammonium di-tert-butylphosphate (30.4 mg, 0.067 mmol) in dimethoxyethane (2 mL) was stirred at 80° C. for 3.5 hours. The reaction mixture was cooled, quenched with brine, and extracted with ethyl acetate (2×). The combined organic layers were dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The residue was purified on a 12 g column using the Biotage Isolera™ One flash system eluted with heptanes/ethyl acetate (2:8 to 1:9) to provide the title compound (35.0 mg, 66%). MS (EST⁺) m/z 823.3 (M+H)⁺.

Example 20C: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 3-(phosphonooxy)propyl Carbonate

A mixture of Example 20B (41.0 mg, 0.050 mmol) and trifluoroacetic acid (0.077 mL, 0.996 mmol) in CH₂Cl₂ (0.5 mL) was stirred for 2 hours. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by reverse-phase HPLC performed on a custom packed YMC TriArt™ C18 Hybrid plastic column (20 μm, 25×70 mm) using a gradient of 5-100% acetonitrile:0.1% aqueous trifluoroacetic acid at a flow rate of 70 mL/minute to provide the title compound (27.8 mg, 78%). ¹H NMR (400 MHz, CD₃OD) δ ppm 7.36 (td, J=8.7, 2.0 Hz, 2H), 6.89 (dt, J=11.0, 3.1 Hz, 2H), 6.79 (ddt, J=8.9, 2.7, 1.3 Hz, 2H), 5.43 (dd, J=9.7, 2.0 Hz, 1H), 4.50-4.36 (m, 4H), 4.23 (ddt, J=10.8, 6.1, 4.6 Hz, 2H), 4.03 (q, J=6.3 Hz, 2H), 2.58 (ddd, J=14.2, 9.3, 2.9 Hz, 1H), 2.47-2.33 (m, 1H), 2.22-1.88 (m, 10H); MS (ESI m/z 711.1 (M+H)⁺.

Example 21: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 4-(phosphonooxy)butanoate (Compound 120) Example 21A: methyl 4-{[bis(benzyloxy)phosphoryl]oxy}butanoate

To a solution of methyl 4-hydroxybutanoate (0.565 g, 4.78 mmol) in 0.45 M tetrazole (15.9 mL, 7.17 mmol) in acetonitrile at 0° C. was added dibenzyl diethylphosphoramidite (1.769 mL, 5.02 mmol) dropwise. The mixture was warmed to room temperature and stirred for 1 hour. The reaction mixture was cooled to 0° C. again and 30% hydrogen peroxide in water (1.44 mL, 14.35 mmol) was added. After being stirred for 30 minutes at 0° C., the reaction mixture was quenched with saturated, aqueous Na₂S₂O₃ and brine, and extracted with ethyl acetate (2×). The combined organic layers were dried over anhydrous MgSO₄, filtered, concentrated under reduced pressure and purified on a 120 g silica gel column using the Biotage Isolera™ One flash system eluted with heptanes/ethyl acetate (5:5 to 4:6) to provide the title compound (0.660 g, 37%). MS (EST⁺) m/z 379.1 (M+H)⁺.

Example 21B: 4-{[bis(benzyloxy)phosphoryl]oxy}butanoic Acid

A mixture of Example 21A (0.575 g, 1.520 mmol) in tetrahydrofuran (7 mL) and methanol (5 mL) was treated with a solution of lithium hydroxide (0.109 g, 4.56 mmol) in water (3.5 mL). The mixture was stirred for 3.5 hour and most of the solvent had evaporated. The residue was diluted with water (2 mL), treated with 5% citric acid until pH=4, and extracted with ethyl acetate (2×). The combined organic layers were dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The residue was purified on a 40 g silica gel column using the Biotage Isolera™ One flash system eluted with heptanes/ethyl acetate (2:8 to 1:9) to provide the title compound (36.5 mg, 6.6%). MS (ESI⁺) m/z 365.5 (M+H)⁺.

Example 21C: (2 S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 4-{[bis(benzyloxy)phosphoryl]oxy}butanoate

To a solution of the product of Example 1L (0.045 g, 0.085 mmol), 4-dimethylaminopyridine (0.011 g, 0.094 mmol), and Example 21B (0.034 g, 0.094 mmol) in N,N-dimethylformamide (1.2 mL) was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC, 0.024 g, 0.128 mmol). The mixture was stirred overnight and additional EDC (15 mg) was added. The reaction mixture was stirred for an additional 24 hours. The reaction mixture was then quenched with brine and extracted with ethyl acetate (2×). The combined organic layers were washed with brine, dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The residue was purified on a 12 g silica gel column using the Biotage Isolera™ One flash system eluted with heptanes/ethyl acetate (2:8 to 1:9) to provide the title compound mixed with 30% of Example 1L (53.0 mg). This material was carried into the next step without further purifications. MS (ESI⁺) m/z 875.3 (M+H)⁺.

Example 21D: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 4-(phosphonooxy)butanoate

A mixture of Example 21C (50.0 mg) and trifluoroacetic acid (0.132 mL, 1.713 mmol) in CH₂Cl₂ (0.4 mL) was stirred at room temperature for 16 hours, then at 35° C. for 4 hours. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by reverse-phase HPLC performed on a custom packed YMC TriArt™ C18 Hybrid plastic column (20 μm, 25×70 mm) using a gradient of 5-100% acetonitrile:0.1% aqueous trifluoroacetic acid at a flow rate of 70 mL/minute to provide the title compound (25.5 mg). ¹H NMR (400 MHz, CD₃OD) δ ppm 7.36 (t, J=8.7 Hz, 2H), 6.90 (dd, J=11.0, 2.8 Hz, 2H), 6.79 (dddd, J=9.0, 4.1, 2.9, 1.3 Hz, 2H), 5.57-5.48 (m, 1H), 4.49-4.33 (m, 4H), 3.99 (q, J=6.3 Hz, 2H), 2.54 (ddd, J=14.1, 9.3, 3.0 Hz, 1H), 2.49-2.41 (m, 2H), 2.41-2.29 (m, 1H), 2.26-1.82 (m, 10H); MS (ESI⁺) m/z 695.2 (M+H)⁺.

Example 22: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 2-(phosphonooxy)ethyl Carbonate (Compound 121) Example 22A: dibenzyl 2-hydroxyethyl Phosphate

To a solution of 2-((tert-butyldimethylsilyl)oxy)ethanol (0.60 g, 3.38 mmol) in a 0.5 M acetonitrile solution of 1H-tetrazole (11.2 mL, 5.04 mmol) at 0° C. was slowly added dibenzyl diisopropylphosphoramidite (1.36 mL, 4.06 mmol). The mixture was allowed to warm to ambient temperature and was stirred for 2 hours. The material was cooled to 0° C., quenched with 30% aqueous hydrogen peroxide (1.05 mL, 10.28 mmol) and allowed to warm to ambient temperature. The mixture was stirred for 15 minutes, diluted with methanol (10 mL), acidified with concentrated HCl (1.4 mL, 16.8 mmol) and stirred for 20 minutes. The reaction was neutralized with concentrated NH₄OH (2.2 mL, 17.5 mmol) and then concentrated under reduced pressure to remove most volatiles. The residue was diluted with ethyl acetate, washed with water and brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified on silica gel (50-100% ethyl acetate/heptanes) to give the title compound (0.853 g, 2.65 mmol, 78% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.48-7.25 (m, 10H), 5.04 (d, J=7.8 Hz, 4H), 4.92 (t, J=5.6 Hz, 1H), 3.97 (ddd, J=7.3, 5.4, 4.5 Hz, 2H), 3.56 (tdd, J=5.7, 4.4, 1.3 Hz, 2H); LC/MS (APCI⁺) m/z 323 (M+H)⁺.

Example 22B: 2-{[bis(benzyloxy)phosphoryl]oxy}ethyl (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Carbonate

A solution of Example 22A (0.0414 g, 0.128 mmol), Example 9A (0.0754 g, 0.112 mmol), N,N-dimethylpyridin-4-amine (0.016 g, 0.133 mmol) and triethylamine (0.017 mL, 0.122 mmol) in tetrahydrofuran (0.55 mL) was stirred at ambient temperature for 24 hours. The mixture was diluted with ethyl acetate, washed with 1 N HCl, 1 N NaOH, and brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified on silica gel (25% methyl tert-butyl ether/dichloromethane) to give the title compound (0.056 g, 0.064 mmol, 56.5% yield). ¹H NMR (501 MHz, DMSO-d₆) (5 ppm 7.65 (s, 1H), 7.62 (s, 1H), 7.46 (t, J=8.9 Hz, 1H), 7.42 (t, J=8.8 Hz, 1H), 7.39-7.30 (m, 10H), 7.01 (dd, J=11.4, 2.9 Hz, 1H), 6.96 (dd, J=11.4, 2.8 Hz, 1H), 6.80 (ddd, J=9.0, 3.0, 1.1 Hz, 1H), 6.76 (ddd, J=9.0, 2.9, 1.1 Hz, 1H), 5.29 (dd, J=9.5, 2.2 Hz, 1H), 5.03 (dd, J=8.1, 3.7 Hz, 4H), 4.45-4.38 (m, 4H), 4.32-4.24 (m, 1H), 4.24-4.15 (m, 3H), 2.43 (ddd, J=14.3, 9.2, 2.8 Hz, 1H), 2.32-2.22 (m, 1H), 1.99-1.92 (m, 2H), 1.92-1.73 (m, 6H); LC/MS (APCI⁺) m/z 877 (M+H)⁺.

Example 22C: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 2-(phosphonooxy)ethyl Carbonate

A solution of Example 22B (0.056 g, 0.064 mmol) in CH₂Cl₂ (0.30 mL) and trifluoroacetic acid (0.15 mL, 0.064 mmol) was stirred at ambient temperature for 4 hours then was heated to 35° C. and allowed to stir for 22 hours. The mixture was concentrated under reduced pressure, and the residue was purified by preparative LC (YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, 140 mL/minute, 0-70% gradient of acetonitrile in 25 mM aqueous NH₄HCO₃ over 10 minutes) to give the title compound (0.028 g, 0.040 mmol, 63% yield). ¹H NMR (400 MHz, DMSO-d₆) (5 ppm 8.39 (s, 1H), 7.69 (s, 1H), 7.45 (dt, J=17.6, 8.9 Hz, 2H), 7.14 (s, 2H), 7.02 (ddd, J=11.4, 5.0, 2.8 Hz, 2H), 6.81 (dd, J=9.0, 2.8 Hz, 2H), 5.22 (d, J=9.1 Hz, 1H), 4.57-4.44 (m, 2H), 4.43 (s, 2H), 4.31-4.18 (m, 1H), 4.17-4.10 (m, 1H), 3.99-3.77 (m, 2H), 2.47-2.37 (m, 1H), 2.34-2.21 (m, 1H), 2.02-1.72 (m, 8H); MS (ESI) m/z 694.8 (M−H)⁻.

Example 23: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 3,3-dimethyl-4-(phosphonooxy)butanoate (Compound 122) Example 23A: tert-butyl 4-{[bis(benzyloxy)phosphoryl]oxy}-3,3-dimethylbutanoate

To a solution of 3,3-dimethyldihydrofuran-2,5-dione (2.0 g, 15.61 mmol) in tetrahydrofuran (75 mL) at 0° C. was added a 1.0 M solution of potassium 2-methylpropan-2-olate in tetrahydrofuran (16.4 mL, 16.4 mmol) dropwise. The suspension was then warmed to ambient temperature and allowed to stir overnight. The reaction mixture was quenched with saturated ammonium chloride solution and then acidified to pH 3 with 3 N HCl. The aqueous phase was extracted with ethyl acetate (2×100 mL). The combined organic layers was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under vacuum to give 2.38 g of crude as light yellow oil. To a solution of this material (1.0 g, 4.94 mmol) in tetrahydrofuran (30 mL) at 0° C. was added a 1.0 M tetrahydrofuran solution of borane tetrahydrofuran complex (9.89 mL, 9.89 mmol) dropwise. The mixture was stirred at ambient temperature for 3 hours, and methanol was added. The resulting solution was stirred overnight. The solvent was removed under reduced pressure to give intermediate, tert-butyl 4-hydroxy-3,3-dimethylbutanoate which was used without further purification.

To a solution of tert-butyl 4-hydroxy-3,3-dimethylbutanoate (0.35 g, 1.859 mmol), N-ethyl-N-isopropylpropan-2-amine (0.974 mL, 5.58 mmol) and tetrabenzyl diphosphate (2.403 g, 4.46 mmol) in CH₂Cl₂ (7.5 mL) was added tetra-tert-butoxy titanium (0.127 g, 0.372 mmol) at ambient temperature, and the solution was stirred for 2 hours. The solvent was removed under reduced pressure, and the residue was purified on silica gel using a Biotage Isolera™ One flash system eluted with ethyl acetate/heptanes (0-50%) to give 0.25 g of the title compound. ¹H NMR (400 MHz, DMSO-d₆) (5 ppm 7.44-7.27 (m, 10H), 5.04 (dd, J=8.2, 3.8 Hz, 4H), 3.76 (d, J=4.6 Hz, 2H), 2.10 (s, 2H), 1.38 (s, 9H), 0.92 (s, 6H).

Example 23B: 4-{[bis(benzyloxy)phosphoryl]oxy}-3,3-dimethylbutanoic Acid

To a solution of Example 23A (390 mg, 0.722 mmol) in dichloromethane (5.0 mL) at room temperature was added trifluoroacetic acid (0.56 mL, 7.22 mmol), and the mixture was stirred for 3 hours. Additional trifluoroacetic acid (3.61 mmol) was added, and the mixture was stirred at 40° C. for 2 hours. The trifluoroacetic acid and CH₂Cl₂ were removed under high vacuum, and the residue was purified by HPLC (Phenomenex® Luna® C18(2) 10 μm 100 Å AXIA™ column (250 mm×50 mm). A 20-90% gradient of acetonitrile (A) and 0.1% trifluoroacetic acid in water (B) was used over 25 minutes, at a flow rate of 50 mL/minute) to give 250 mg of the title compound. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.51-7.10 (m, 10H), 5.04 (d, J=8.3 Hz, 4H), 3.81 (d, J=4.6 Hz, 2H), 2.16 (s, 2H), 0.94 (s, 6H); MS (ESI+) m/z 393.2 (M+H)⁺.

Example 23C: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 4-{[bis(benzyloxy)phosphoryl]oxy}-3,3-dimethylbutanoate

To a solution of the product of Example 1L (320 mg, 0.605 mmol), Example 23B (226 mg, 0.576 mmol) and 4-dimethylaminopyridine (91 mg, 0.749 mmol) in N,N-dimethylformamide (3.0 mL) was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (199 mg, 1.04 mmol), and the mixture was stirred at room temperature overnight. N,N-Dimethylformamide was removed under high vacuum, and the residue was purified by HPLC (Phenomenex® Luna® C18(2) 10 μm 100 Å AXIA™ column (250 mm×50 mm). A 40-100% gradient of acetonitrile (A) and 0.1% trifluoroacetic acid in water (B) was used over 25 minutes, at a flow rate of 50 mL/minute) to give 300 mg of the title compound. ¹H NMR (400 MHz, DMSO-d₆) (5 ppm 7.64-7.21 (m, 14H), 7.10-6.91 (m, 2H), 6.77 (dtdd, J=11.8, 8.9, 2.9, 1.2 Hz, 2H), 5.12-4.96 (m, 4H), 4.50-4.31 (m, 6H), 3.73 (d, J=4.7 Hz, 2H), 2.33-2.11 (m, 2H), 2.11-1.70 (m, 8H), 0.88 (s, 6H); MS (ESI+) m/z 903.3 (M+H)⁺.

Example 23D: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 3,3-dimethyl-4-(phosphonooxy)butanoate

A mixture of Example 23C (120 mg, 0.133 mmol), Pd/C (22.6 mg, 10.6 μmol), and 4 N HCl solution in dioxane (0.066 mL, 0.266 mmol) in tetrahydrofuran (5.0 mL) in a 20 mL Barnstead Hast C vessel was stirred and hydrogenated with 50 psi of hydrogen at 25° C. for 0.5 hour. After the reaction was complete, the mixture was filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by preparative HPLC [custom packed YMC TriArt™ C18 Hybrid 20 μm plastic column, 25×70 mm, flow rate 70 mL/minute, 0-100% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid)] to give 49 mg of the title compound as a white solid. ¹H NMR (400 MHz, DMSO-d₆) (5 ppm 7.58 (d, J=2.4 Hz, 2H), 7.43 (td, J 8.9, 4.4 Hz, 2H), 7.03-6.91 (m, 2H), 6.76 (ddd, J=8.9, 5.5, 2.9 Hz, 2H), 5.30 (dd, J=9.4, 2.2 Hz, 1H), 4.39 (m, 4H), 3.57 (s, 2H), 2.38 (ddd, J=14.2, 9.4, 2.7 Hz, 1H), 2.25 (dd, J=13.2, 3.9 Hz, 1H), 2.20 (s, 2H), 2.01-1.71 (m, 8H), 0.92 (s, 6H); MS (ESI+) m/z 723.2 (M+H)⁺.

Example 24: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Methyl Carbonate (Compound 123)

The product of Example 17D (20 mg, 0.034 mmol) was dissolved in methanol (1.0 mL) and stirred at ambient temperature for 10 minutes. The resulting solution was concentrated under reduced pressure, and the residue was purified by preparative HPLC [YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, flow rate 140 mL/minute, 5-100% gradient of acetonitrile in buffer (0.025 M aqueous ammonium bicarbonate, adjusted to pH 10 with ammonium hydroxide)] to give the title compound (15 mg, 0.026 mmol, 76% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.67 (s, 1H), 7.64 (s, 1H), 7.50-7.42 (m, 2H), 7.05-6.93 (m, 2H), 6.84-6.73 (m, 2H), 5.31-5.22 (m, 1H), 4.45 (s, 2H), 4.44 (s, 2H), 3.68 (s, 3H), 2.44-2.35 (m, 1H), 2.34-2.21 (m, 1H), 2.08-1.73 (m, 8H); MS (ESI⁺) m/z 587 (M+H)⁺.

Example 25: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl dimethylcarbamate (Compound 124)

The product of Example 17D (20 mg, 0.034 mmol) was added to dimethylamine (2.0 M in tetrahydrofuran, 0.5 mL). The resulting mixture was stirred at ambient temperature for 10 minutes and then concentrated under reduced pressure. The residue was purified by preparative HPLC [YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, flow rate 140 mL/minute, 5-100% gradient of acetonitrile in buffer (0.025 M aqueous ammonium bicarbonate, adjusted to pH 10 with ammonium hydroxide)] to give the title compound (18 mg, 0.03 mmol, 89% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.68 (s, 1H), 7.60 (s, 1H), 7.50-7.42 (m, 2H), 7.05-6.96 (m, 2H), 6.82-6.76 (m, 2H), 5.09-5.05 (m, 1H), 4.48-4.38 (m, 4H), 2.82 (s, 3H), 2.80 (s, 3H), 2.41-2.29 (m, 1H), 2.17-1.71 (m, 9H); MS (ESI⁺) m/z 600 (M+H)⁺.

Example 26: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Hydrogen Sulfate (Compound 125)

To a suspension of Example 1L (0.048 g, 0.090 mmol) in CH₂Cl₂ (0.50 mL) at ambient temperature was added sulfurochloridic acid (8 μL, 0.120 mmol), and the mixture was stirred for 2.5 hours. The mixture was concentrated under reduced pressure, and the residue was purified on silica gel (12% methanol/dichloromethane) to give the title compound (0.040 g, 0.065 mmol, 72% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.10 (s, 1H), 7.56 (s, 1H), 7.47 (td, J=8.9, 4.0 Hz, 2H), 7.10 (dd, J=11.3, 2.8 Hz, 1H), 7.03 (dd, J=11.4, 2.8 Hz, 1H), 6.94-6.87 (m, 1H), 6.81 (ddd, J=8.9, 2.9, 1.2 Hz, 1H), 4.44 (s, 2H), 4.40-4.32 (m, 3H), 2.72 (ddd, J=13.0, 11.0, 4.0 Hz, 1H), 2.39 (ddd, J=12.9, 9.6, 3.0 Hz, 2H), 2.05-1.73 (m, 5H), 1.73-1.61 (m, 1H), 1.49-1.37 (m, 1H); MS (ESI) m/z 607.1 (M−H)⁻.

Example 27: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Sulfamate (Compound 126)

To a solution of the product of Example 1L (0.075 g, 0.14 mmol) in dimethylacetamide (1.5 mL) was added sulfamoyl chloride (0.094 g, 0.81 mmol) at ambient temperature. The reaction mixture stirred for 16 hours and then was purified by preparative HPLC [Waters XBridge™ C18 5 μm OBD column, 50×100 mm, flow rate 90 mL/minute, 5-100% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid)] to give the title compound (0.056 g, 0.092 mmol, 65% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.68 (s, 1H), 7.54-7.42 (m, 5H), 7.03 (ddd, J=11.3, 2.9, 1.9 Hz, 2H), 6.82 (dddd, J=8.9, 4.8, 2.8, 1.2 Hz, 2H), 5.04 (d, J=9.0 Hz, 1H), 4.44 (d, J=7.0 Hz, 4H), 2.22 (dd, J=25.9, 11.6 Hz, 3H), 2.09-1.91 (m, 3H), 1.83 (t, J=10.5 Hz, 4H); MS (ESI⁺) m/z 608 (M+H)⁺.

Example 28: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 5-(phosphonooxy)pentanoate (Compound 127) Example 28A: methyl 5-[(di-tert-butoxyphosphoryl)oxy]pentanoate

A mixture of methyl 5-bromopentanoate (1.00 mL, 6.99 mmol) and tetra-N-butylammonium di-tert-butylphosphate (3.31 g, 7.34 mmol) in dimethoxyethane (20 mL) was stirred at 80° C. for 3 hours. The reaction was cooled to ambient temperature and was quenched with brine and extracted with ethyl acetate (2×). The combined organic layers were dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The residue was purified on an 80 g silica gel column using the Biotage Isolera™ One flash system eluted with heptanes/ethyl acetate (3:7 to 2:8) to provide the title compound (1.68 g, 74%). MS (DCI⁺) m/z 325.1 (M+H)⁺.

Example 28B: 5-[(di-tert-butoxyphosphoryl)oxy]pentanoic Acid

A solution of Example 28A (1.40 g, 4.32 mmol) in tetrahydrofuran (15 mL) and methanol (10 mL) was treated with a solution of lithium hydroxide monohydrate (0.453 g, 10.8 mmol) in water (8 mL). The mixture was stirred for 3 hours, and then most solvents were evaporated under reduced pressure. The residue was diluted with 2 mL of water and acidified with 5% citric acid until pH 5. The resulting emulsion was extracted with ethyl acetate (2×). The combined organic layers were washed with brine, dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure to provide the title compound (1.12 g, 84%). MS (DCI⁺) m/z 311.1 (M+H)⁺.

Example 28C: (28)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 5-[(di-tert-butoxyphosphoryl)oxy]pentanoate

To a solution of the product of Example 1L (0.300 g, 0.567 mmol), 4-dimethylaminopyridine (0.076 g, 0.623 mmol), and Example 28B (0.211 g, 0.680 mmol) in N,N-dimethylformamide (8 mL) was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC, 0.163 g, 0.850 mmol). The mixture was stirred overnight, then was quenched with brine, and extracted with ethyl acetate (2×). The combined organic layers were washed with brine, dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The residue was purified on a 40 g silica gel column using the Biotage Isolera™ One flash system eluted with heptanes/ethyl acetate (3:7 to 1:9) to provide the title compound (0.360 g, 77%). MS (ESI⁺) m/z 820.7 (M+H)⁺.

Example 28D: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl 5-(phosphonooxy)pentanoate

A mixture of Example 28C (0.355 g, 0.432 mmol) and trifluoroacetic acid (0.5 mL, 6.48 mmol) in CH₂Cl₂ (4 mL) was stirred for 3 hours. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by reverse-phase HPLC performed on a YMC TriArt™ C18 Hybrid column (5 μm, 50×100 mm) using a gradient of 3-100% acetonitrile:0.1% aqueous trifluoroacetic acid at a flow rate of 140 mL/minute to provide the title compound (0.206 g, 67%). ¹H NMR (400 MHz, CD₃OD) δ ppm 7.36 (td, J=8.7, 2.6 Hz, 2H), 6.90 (ddd, J=11.0, 2.9, 1.0 Hz, 2H), 6.79 (dddd, J=8.9, 4.0, 2.8, 1.2 Hz, 2H), 5.52 (dt, J=9.3, 1.8 Hz, 1H), 4.48-4.32 (m, 4H), 4.04-3.89 (m, 2H), 2.54 (ddd, J=14.2, 9.4, 3.1 Hz, 1H), 2.34 (qd, J=6.9, 2.6 Hz, 3H), 2.27-1.87 (m, 8H), 1.78-1.59 (m, 4H); MS (ESI m/z 709.1 (M+H)⁺.

Example 29: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl pyridine-3-carboxylate (Compound 128)

To a suspension of Example 1L (0.100 g, 0.19 mmol), triethylamine (0.034 mL, 0.244 mmol) and N,N-dimethylpyridin-4-amine (0.0234 g, 0.192 mmol) in CH₂Cl₂ (1 mL) was added nicotinoyl chloride (0.0299 g, 0.211 mmol), and the mixture was stirred overnight. Additional nicotinoyl chloride (0.0299 g, 0.211 mmol) and triethylamine (0.034 mL, 0.244 mmol) were added, and the mixture was stirred for 90 minutes. The reaction mixture was diluted with ethyl acetate, quenched with 1 N HCl (0.7 mL), washed with water and brine, and dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified on silica gel (35% ethyl acetate/dichloromethane) to give the title compound (0.0910 g, 0.143 mmol, 76% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.09 (dd, J=2.2, 0.9 Hz, 1H), 8.83 (dd, J=4.8, 1.7 Hz, 1H), 8.26 (dt, J=7.9, 1.9 Hz, 1H), 7.76 (s, 1H), 7.65 (s, 1H), 7.57 (ddd, J=7.9, 4.9, 0.9 Hz, 1H), 7.46 (t, J=8.9 Hz, 1H), 7.21 (t, J=8.8 Hz, 1H), 7.01 (dd, J=11.4, 2.9 Hz, 1H), 6.85 (dd, J=11.3, 2.9 Hz, 1H), 6.79 (ddd, J=9.0, 2.9, 1.2 Hz, 1H), 6.65 (ddd, J=8.9, 2.8, 1.2 Hz, 1H), 5.58 (d, J=9.1 Hz, 1H), 4.43 (s, 2H), 4.41 (s, 2H), 2.52 (buried), 2.44-2.34 (m, 1H), 2.20 (t, J=11.5 Hz, 1H), 2.10 (t, J=12.3 Hz, 1H), 2.03-1.74 (m, 6H); MS (ESI) m/z 632.1 (M−H)⁻.

Example 30: 3-[({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)carbonyl]-1-methylpyridin-1-ium iodide (Compound 129)

A solution of Example 29 (0.075 g, 0.119 mmol) and iodomethane (37 μL, 0.594 mmol) in acetone (1.0 mL) was stirred at 60° C. overnight. The mixture was then concentrated under reduced pressure and triturated with methyl tert-butyl ether (˜1 mL). The resulting solids were isolated via filtration and dried under vacuum to give the title compound (0.090 g, 0.116 mmol, 98% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.45 (s, 1H), 9.21 (d, J=6.1 Hz, 1H), 8.94 (dt, J=8.2, 1.6 Hz, 1H), 8.27 (dd, J=8.1, 6.0 Hz, 1H), 7.69 (d, J=6.6 Hz, 2H), 7.46 (t, J=8.9 Hz, 1H), 7.33 (t, J=8.9 Hz, 1H), 7.00 (dd, J=11.4, 2.9 Hz, 1H), 6.89 (dd, J=11.4, 2.9 Hz, 1H), 6.79 (ddd, J=9.0, 2.9, 1.2 Hz, 1H), 6.69 (ddd, J=9.0, 2.9, 1.2 Hz, 1H), 5.62 (d, J=9.2 Hz, 1H), 4.44 (t, J=3.5 Hz, 7H), 2.60-2.52 (m, 1H), 2.35 (dd, J=20.9, 11.1 Hz, 2H), 2.24-2.13 (m, 1H), 2.08 (m, with acetone impurity), 2.00-1.77 (m, 5H); MS (ESI m/z 648.3 (M)⁺.

Example 31: (2R)-2-{[({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)carbonyl]amino}pentanedioic acid (Compound 130)

To a solution of the product of Example 9A (0.08 g, 0.12 mmol) and (R)-2-aminopentanedioic acid (0.070 g, 0.48 mmol) in tetrahydrofuran (0.8 mL) was added triethylamine (0.03 mL, 0.2 mmol). This reaction mixture stirred at ambient temperature for 22 hours. The mixture was then concentrated under reduced pressure, diluted with N,N-dimethylformamide (1 mL), filtered and purified by preparative HPLC [Waters XBridge™ C18 5 μm OBD column, 50×100 mm, flow rate 90 mL/minute, 5-100% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid)] to afford the title compound (0.015 g, 0.021 mmol, 18% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.62 (d, J=9.9 Hz, 1H), 7.55 (t, J=10.7 Hz, 1H), 7.46 (dt, J=13.2, 9.1 Hz, 2H), 7.07-6.98 (m, 2H), 6.80 (s, 2H), 6.54-6.49 (m, 1H), 5.15 (d, J=9.1 Hz, 1H), 4.43 (d, J=4.3 Hz, 3H), 4.36 (d, J=14.7 Hz, 1H), 4.00 (s, 1H), 2.34-2.26 (m, 2H), 2.05 (s, 5H), 1.92 (d, J=13.8 Hz, 3H), 1.82 (s, 4H); MS (ESI⁺) m/z 702 (M+H)⁺.

Example 32: ({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)methyl Dihydrogen Phosphate (Compound 131) Example 32A: N,N′-{(2S)-2-[(methylsulfanyl)methoxy]bicyclo[2.2.2]octane-1,4-diyl}bis[2-(4-chloro-3-fluorophenoxy)acetamide]

A suspension of sodium hydride (6.80 g, 170 mmol) in tetrahydrofuran (200 mL) was stirred at ambient temperature, and iodine (4.79 g, 18.89 mmol) was added as a solid in one portion. The product of Example 1L (20.0 g, 37.8 mmol) was added, and the mixture was heated to 40-45° C. for 20 minutes. The mixture was then cooled to <5° C., and chloromethyl methyl sulfide (6.24 mL, 76 mmol) was added, maintaining the temperature at <5° C. The mixture was stirred at that temperature for 6 hours then was poured into a vigorously stirred mixture of ethyl acetate (200 mL) and saturated aqueous NH₄Cl (200 mL) pre-cooled to 5° C. After warming to ambient temperature, the layers were separated, and the organic layer was washed with brine (40 mL) and dried over anhydrous Na₂SO₄. The material was concentrated under reduced pressure, and the residue was purified on silica gel (750 g Teledyne ISCO RediSep Gold® cartridge, 120 mL/minute, 0-20% methyl tert-butyl ether/dichloromethane gradient over 60 minutes) to give the title compound (14.5 g, 24.60 mmol, 65.1% yield). ¹H NMR (501 MHz, DMSO-d₆) δ ppm 7.58 (s, 1H), 7.48 (td, J=8.9, 4.3 Hz, 2H), 7.30 (s, 1H), 7.03 (ddd, J=11.5, 7.6, 2.8 Hz, 2H), 6.81 (dddd, J=8.8, 4.2, 2.9, 1.2 Hz, 2H), 4.64 (d, J=11.4 Hz, 1H), 4.60 (d, J=11.3 Hz, 1H), 4.45 (s, 2H), 4.43 (s, 2H), 4.29-4.24 (m, 1H), 2.28 (ddd, J=13.6, 9.1, 2.6 Hz, 1H), 2.21-2.12 (m, 1H), 2.06 (s, 3H), 2.01-1.93 (m, 1H), 1.93-1.73 (m, 7H); LC/MS (APCI⁺) m/z 591 (M+H)⁺.

Example 32B: ({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)methyl Disodium Phosphate

A suspension of Example 32A (16.9 g, 28.7 mmol) and crystalline H₃PO₄ (19.48 g, 199 mmol) in tetrahydrofuran (170 mL) was stirred with activated 5 Å molecular sieves (85 g) at ambient temperature for 5 minutes. The mixture was cooled to 0° C., and solid N-iodosuccinimide (9.69 g, 43.1 mmol) was added at once. The mixture was stirred for 35 minutes then was filtered through diatomaceous earth with ethyl acetate (200 mL) and quenched into saturated, aqueous NaHCO₃ (580 mL) and saturated, aqueous Na₂S₂O₃ (75 mL). The layers were separated, and the organic fraction was washed with saturated, aqueous NaHCO₃ (100 mL). The aqueous layers were back-extracted with ethyl acetate (100 mL) containing toluene (50 mL). The combined organic layers were concentrated to give 30 g of light yellow solid, which was triturated with methyl tert-butyl ether (3 volumes). The solid was collected by filtration, and the solids were rinsed with methyl tert-butyl ether and ethyl acetate The solids was partially vacuum dried to give 23 g of white solid, which was dissolved in boiling ethanol (60 mL) and hot-filtered. The residual solids were rinsed with ethanol (10 mL). The filtrate was concentrated under reduced pressure to give 20 g of solid, which was precipitated from water (6 mL) and isopropyl alcohol (60 mL). The precipitate was collected by filtration, and the solids were rinsed with isopropyl alcohol, azeotroped with toluene (50 mL) and vacuum dried to give a white powder. This material was slurried with water (30 mL), the solids were collected by filtration, and the collected solids were washed with water (5 mL). The wet solid was then dissolved in water (65 mL) at 80° C. and allowed to slowly cool to 40° C. The material was then cooled to 10° C., and the precipitate was collected by filtration. The solids were rinsed with cold water (15 mL) and dried under vacuum to give the title compound (12.4 g, 16.4 mmol, 57% yield, 4.56 weight % water by KF). ¹H NMR (400 MHz, methanol-d₄) δ ppm 7.39-7.33 (m, 1H), 7.30 (d, J=8.8 Hz, 1H), 6.94 (dd, J=11.2, 2.9 Hz, 1H), 6.90 (dd, J=11.0, 2.8 Hz, 1H), 6.85 (ddd, J=8.9, 2.9, 1.2 Hz, 1H), 6.79 (ddd, J=9.0, 2.9, 1.3 Hz, 1H), 5.21 (dd, J=7.3, 5.7 Hz, 1H), 4.97 (dd, J=15.8, 5.8 Hz, 1H), 4.63 (dd, J=2.6 Hz, 2H), 4.41 (s, 2H), 4.30 (d, J=9.2 Hz, 1H), 2.75 (ddd, J=13.1, 11.1, 4.3 Hz, 1H), 2.53-2.37 (m, 2H), 2.13-1.85 (m, 6H), 1.62 (dddd, J=13.2, 10.8, 5.7, 2.5 Hz, 1H); MS (ESI) m/z 636.9 (M−H)⁻.

Example 33: (2R)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Acetate (Compound 132)

The first eluting compound from the chiral separation described in Example 1M was the title compound (3.23 g, 5.65 mmol, 38% yield). ¹H NMR (501 MHz, DMSO-d₆) δ ppm 7.60 (s, 1H), 7.55 (s, 1H), 7.47 (td, J=8.9, 2.0 Hz, 2H), 7.00 (ddd, J=16.6, 11.4, 2.9 Hz, 2H), 6.79 (dddd, J=8.6, 7.1, 2.9, 1.1 Hz, 2H), 5.28 (dd, J=9.5, 2.4 Hz, 1H), 4.48-4.39 (m, 4H), 2.39 (ddd, J=14.1, 9.4, 2.9 Hz, 1H), 2.21 (dd, J=11.6, 7.6 Hz, 1H), 2.02 (td, J=8.4, 2.8 Hz, 1H), 1.98 (s, 3H), 1.93 (dd, J=9.1, 6.9 Hz, 2H), 1.89-1.76 (m, 5H); MS (EST⁺) m/z 570.8 (M+H)⁺.

Example 34: (2R)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (methylamino)acetate (Compound 133) Example 34A: N,N′-[(2R)-2-hydroxybicyclo[2.2.2]octane-1,4-diyl]bis[2-(4-chloro-3-fluorophenoxy)acetamide]

The title compound was isolated using the chiral preparative SFC described in Example 1L as the first peak eluted off the column. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.53-7.43 (m, 3H), 7.28 (s, 1H), 7.04 (ddd, J=12.3, 11.5, 2.9 Hz, 2H), 6.82 (tdd, J=9.0, 2.9, 1.2 Hz, 2H), 5.10 (s, 1H), 4.45 (d, J=13.5 Hz, 4H), 4.04 (dd, J=9.6, 3.1 Hz, 1H), 2.27 (ddd, J=12.2, 9.4, 2.2 Hz, 1H), 2.07 (ddd, J=12.2, 10.4, 4.7 Hz, 1H), 1.96-1.72 (m, 8H); MS (EST⁺) m/z 529.1 (M+H)⁺.

Example 34B (2R)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (methylamino)acetate

A suspension of the product of Example 34A (50 mg, 0.094 mmol) in N,N-dimethylformamide (1 tut) and N,N-diisopropylethylamine (0.041 mL, 0.236 mmol) was treated with 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxid hexafluorophosphate (40 mg, 0.104 mmol) and with 2-((tert-butyloxycarbonyl)(methyl)amino)acetic acid (27 mg, 0.142 mmol). The reaction mixture was stirred at 25° C. for 20 hours and then was concentrated. Trifluoroacetic acid (1 mL) was added. The mixture was allowed to stir for 10 minutes and then was concentrated under reduced pressure. The residue was purified by HPLC (Phenomenex® Luna® C18(2) 5 μm 100 Å AXIA™ column 250 mm×21.2 mm, flow rate 25 mL/minute, 0-60% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid in water)) to give the title compound as a trifluoroacetic acid salt (45 mg, 0.063 mmol, 67% yield). ¹H NMR (500 MHz, DMSO-do) 6 ppm 8.90 (s, 1H), 8.84 (s, 1H), 7.65 (s, 1H), 7.51 (s, 1H), 7.45 (id, 8.9, 4.8 Hz, 2H), 6.98 (dd. 11.4, 2.9 Hz, 2H), 6.76 (ddt, J=8.9, 2.8, 1.4 Hz, 2H), 5.50 (dtf 9.6, 2.1 Hz, 1H), 4.47-4.34 (in. 4H), 4.03-3.82 (m, 2H), 2.45-2.35 (m, 1H), 2.31-2.22 (m, 1H), 2.12-2.01 (m, 1H), 1.96 (td, J=13.4, 11.4, 5.4 Hz, 2H), 1.90-1.76 (m, 4H), 1.79-1.70 (in, 1H); MS (ESI⁺) m/z 601 (M+H)⁺.

Example 35: (2R)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Aminoacetate (Compound 134)

A suspension of the product of Example 34A (50 mg, 0.094 mmol) in N,N-dimethylformamide (1 mL) and N,N-diisopropylethylamine (0.041 mL, 0.236 mmol) was treated with 1-[bis(dimethylamino)methylene.]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (40 mg, 0.104 mmol) and with 2-((tert-butyloxycarbonyl)amino)acetic acid (25 mg, 0.143 mmol). The reaction mixture was stirred at 25° C. for 20 hours and then was concentrated. Trifluoroacetic acid (1 Int) was added to the residue. The mixture was allowed to stir for 10 minutes and then was concentrated under reduced pressure. The residue as purified by HPLC (Phenomenex® Luna® C18(2) 5 μm 100 Å AXIA™ column 250 mm×21.2 mm, flow rate 25 mL/minute, 0-60% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid in water)) to give the title compound as a trifluoroacetic acid salt (35 mg, 0.03 mmol, 42% yield). ¹H NMR. (500 MHz, DMSO-d₆) δ ppm 8.18 (t, J=5.3 Hz, 3H), 7.65 (s, 1H), 7.51-7.39 (m, 3H), 6.97 (dt, J=11.4, 2.6 Hz, 2H), 6.76 (dtd, J=9.1, 3.2, 1.1 Hz, 2H), 5.48 (dd, J=9.7, 2.6 Hz, 1H), 4.47-4.34 (m, 4H), 3.90-3.68 (m, 2H), 2.46-2.35 (m, 1H), 2.34-2.23 (m, 1H), 2.12-2.01 (m, 1H), 2.02-1.92 (m, 2H), 1.88-1.76 (m, 4H), 1.75 (dd, J=9.0, 5.7 Hz, 1H); MS (ESI⁺) m/z 587 (M+H)⁺.

Example 36: (2R)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl (2S)-2-amino-3-methylbutanoate (Compound 135)

A suspension of the product of Example 34A (50 mg, 0.094 mmol) in N,N-dimethylformamide (1 mL) and N,N-diisopropylethylamine (0.041 mL, 0.236 mmol) was treated with 1-[bis(dimethylamino)methyl me]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (40 mg, 0,104 mmol) and with (S)-2-((tert-butyloxycarbonyl)amino)-3-methylbutanoic acid (31 mg, 0,143 mmol). The reaction mixture was stirred at 25° C. for 20 hours and then was concentrated. Trifluoroacetic acid (0.2 mL) was added to the residue. The mixture was allowed to stir for 10 minutes and then was concentrated under reduced pressure. The residue was purified by HPLC (Phenomenex® Luna® C18(2) 5 μm 100 Å AXIA™ column 250 mm×21.2 mm, flow rate 25 mL/minute, 0-60% gradient of acetonitrile in buffer (0.1% trifluoroacetic acid in water)) to give the title compound as a trifluoroacetic acid salt (25 mg, 0.034 mmol, 36% yield). ¹H NMR (500 MHz, DMSO-d₆) ppm 8.91-8.82 (m, 1H), 7.75 (s, 1H), 7.50 (dt, J=25.1, 8.9 Hz, 2H), 7.11 (dd, J=11.3, 2.8 Hz, 1H), 6.99 (dd, J=11.4, 2.9 Hz, 1H), 6.88 (ddd, J=9.0, 2.8, 1.2 Hz, 1H), 6.79 (ddd, J=8.9, 2.8, 1.2 Hz, 1H), 5.34 (dd, J=9.6, 2.1 Hz, 1H), 4.52-4.41 (m, 2H), 4.2.1 (t, J=5.0 Hz, 2H), 2.47-2.38 (m, 1H), 2.34-2.25 (m, 1H), 2.00 (s, 5H), 1.95-1.85 (m, 4H), 1.79 (t, J=11.1 Hz, 1H), 1.72 (dt, J=13.7, 2.5 Hz, 1H); MS (ESI⁺) m/z 629 (M+H)⁺.

Example 37: N,N′-[(2S)-2-(methoxymethoxy)bicyclo[2.2.2]octane-1,4-diyl]bis[2-(4-chloro-3-fluorophenoxy)acetamide](Compound 136)

A suspension of Example 32A (0.0295 g, 0.050 mmol) and crystalline H₃PO₄ (0.035 g, 0.357 mmol) in tetrahydrofuran (0.60 mL) was stirred with activated 5 Å molecular sieves (150 mg) at 0° C. for 5 minutes, and solid N-iodosuccinimide (0.017 g, 0.075 mmol) was added. The mixture was stirred for 5 minutes and allowed to warm to ambient temperature. The reaction was then stirred for 90 minutes, diluted with 1:1 dichloromethane/methanol, quenched with saturated, aqueous Na₂SO₃ (5 drops) and stirred until the red color dissipated. The material was dried over anhydrous Na₂SO₄, filtered through diatomaceous earth with 1:1 dichloromethane/methanol (˜5 mL total) and concentrated. The residue was purified by preparative LC (YMC TriArt™ C18 Hybrid 5 μm column, 50×100 mm, 140 mL/minute, 10-100% gradient of acetonitrile in saturated aqueous carbonic acid buffer over 11 min) to give the title compound (0.0070 g, 0.012 mmol, 24% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.56 (s, 1H), 7.48 (td, J=8.9, 3.0 Hz, 2H), 7.36 (s, 1H), 7.04 (t, J=2.8 Hz, 1H), 7.01 (t, J=2.8 Hz, 1H), 6.82 (dt, J=2.7, 1.3 Hz, 1H), 6.80 (dt, J=2.9, 1.3 Hz, 1H), 4.60 (d, J=6.6 Hz, 1H), 4.52 (d, J=6.7 Hz, 1H), 4.46 (s, 2H), 4.43 (s, 2H), 4.23 (dd, J=9.6, 2.6 Hz, 1H), 3.24 (s, 3H), 2.28 (dd, J=13.0, 10.2 Hz, 1H), 2.20-2.09 (m, 1H), 2.03-1.70 (m, 8H); MS (ESI⁺) m/z 573.0 (M+H)⁺.

Example 38: {[({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)carbonyl]oxy}methyl[4-(phosphonooxy)phenyl]acetate (Compound 137) Example 38A: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Chloromethyl Carbonate

To a suspension of product of Example 1L (0.302 g, 0.570 mmol) in pyridine (0.30 mL, 3.72 mmol) was added chloromethyl carbonochloridate (0.066 mL, 0.742 mmol), and the slurry was stirred for 3 hours. The mixture was then diluted with ethyl acetate, washed with water, back-extracted with ethyl acetate, washed with brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified on silica gel (15% ethyl acetate/dichloromethane) to give the title compound (0.215 g, 0.346 mmol, 61% yield). ¹H NMR (400 MHz, CDCl₃) δ ppm 7.36-7.27 (m, 2H), 6.78-6.70 (m, 2H), 6.68-6.62 (m, 2H), 6.49 (d, J=3.1 Hz, 1H), 6.14 (s, 1H), 5.96-5.86 (m, 1H), 5.74-5.64 (m, 1H), 5.45 (dt, J=9.5, 2.1 Hz, 1H), 4.40-4.27 (m, 4H), 2.70 (ddd, J=14.3, 9.5, 3.1 Hz, 1H), 2.40-2.27 (m, 1H), 2.27-2.08 (m, 4H), 2.08-1.89 (m, 4H); LC/MS (APCI⁺) m/z 621 (M+H)⁺.

Example 38B: (2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl Iodomethyl Carbonate

A suspension of Example 38A (0.136 g, 0.218 mmol) and sodium iodide (0.098 g, 0.654 mmol) in acetone (1.3 mL) was heated to 55° C. for 60 minutes. The mixture was then allowed to cool to ambient temperature, was diluted with ethyl acetate, washed with brine containing some saturated Na₂S₂O₃ (˜0.5 mL) and dried over anhydrous Na₂SO₄. The material was concentrated under reduced pressure to give the title compound (0.145 g, 0.203 mmol, 93% yield) which was used as-is. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.31 (td, J=8.6, 3.2 Hz, 2H), 6.74 (td, J=10.1, 2.9 Hz, 2H), 6.65 (tdd, J=8.6, 2.9, 1.3 Hz, 2H), 6.50 (s, 1H), 6.14 (s, 1H), 5.94 (d, J=5.1 Hz, 1H), 5.89 (d, J=5.1 Hz, 1H), 5.45 (dt, J=9.5, 2.0 Hz, 1H), 4.41-4.26 (m, 4H), 2.70 (ddd, J=14.4, 9.5, 3.1 Hz, 1H), 2.40-2.27 (m, 1H), 2.27-2.07 (m, 4H), 2.07-1.88 (m, 4H); LC/MS (APCI⁺) m/z 713 (M+H)⁺.

Example 38C: (4-{[bis(benzyloxy)phosphoryl]oxy}phenyl)acetic Acid

Methyl 2-(4-hydroxyphenyl)acetate (25 g, 150 mmol) was dissolved in a commercial solution of 1H-tetrazole (0.45 M in acetonitrile, 501 mL, 226 mmol), and the resulting solution was cooled to <5° C. Dibenzyl diisopropylphosphoramidite (79 mL, 181 mmol) was added dropwise over 5 minutes, and the resulting suspension was stirred at 0° C. (ice-water bath) for an additional 60 minutes. Next, 30% aqueous hydrogen peroxide (23.1 mL, 225 mmol) was added dropwise via syringe over 10 minutes, maintaining an internal temperature below 20° C. The flask was removed from the ice-water bath, and the mixture was stirred for an additional 60 minutes. The material was diluted with 500 mL of water and neat lithium hydroxide hydrate (18.94 g, 451 mmol) was added in one portion. The mixture was stirred at ambient temperature for 1 hour, was washed with methyl tert-butyl ether (500 mL, 2×300 mL), and the aqueous layer was acidified to pH=1 with 6 M HCl. The material was then extracted with methyl tert-butyl ether. The combined extracts were washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude material was purified on silica gel (10-100% methyl tert-butyl ether/heptanes over 30 minutes on 2×330 g silica gel columns) to give the title compound as a colorless oil (31.2 g, 75.7 mmol, 50% yield). ¹H NMR (501 MHz, DMSO-d₆) ppm 7.45-7.30 (m, 10H), 7.31-7.21 (m, 2H), 7.17-7.04 (m, 2H), 5.15 (d, J=8.4 Hz, 4H), 3.56 (s, 2H).

Example 38D: {[({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)carbonyl]oxy}methyl (4-{[bis(benzyloxy)phosphoryl]oxy}phenyl)acetate

To a suspension of 38C (0.216 g, 0.525 mmol) in water (1.6 mL) at 0° C. was added 1 M sodium hydroxide (0.56 mL, 0.560 mmol). The mixture was stirred for 1 hour, and a solution of (nitrooxy)silver (0.104 g, 0.612 mmol) in water (0.40 mL) was then added. The reaction mixture was stirred at 0° C. for 45 minutes, and diethyl ether (0.75 mL) was added. Then the mixture was filtered to give crude (2-(4-((bis(benzyloxy)phosphoryl)oxy)phenyl)acetoxy)silver (0.203 g, 0.390 mmol, 74% yield) as a grey solid, which was used as-is without characterization.

A suspension of the above material (0.106 g, 0.203 mmol) and Example 38B (0.145 g, 0.203 mmol) in acetonitrile (1.0 mL) was stirred for 20 hours. The mixture as diluted with ethyl acetate, filtered through diatomaceous earth, and concentrated under reduced pressure. The residue was purified on silica gel (15-20% ethyl acetate/dichloromethane) to give the title compound (0.108 g, 0.108 mmol, 53% yield). ¹H NMR (501 MHz, DMSO-d₆) δ ppm 7.69 (s, 1H), 7.66 (s, 1H), 7.45 (dt, J=9.6, 8.9 Hz, 2H), 7.41-7.32 (m, 10H), 7.29-7.23 (m, 2H), 7.15-7.09 (m, 2H), 7.00 (ddd, J=18.8, 11.4, 2.8 Hz, 2H), 6.78 (dddd, J=14.4, 9.0, 2.9, 1.1 Hz, 2H), 5.72 (h, J=5.2 Hz, 2H), 5.37 (dd, J=9.5, 2.3 Hz, 1H), 5.14 (d, J=8.4 Hz, 4H), 4.48-4.40 (m, 4H), 3.76 (d, J=1.3 Hz, 2H), 2.44 (ddd, J=14.4, 9.4, 2.7 Hz, 1H), 2.37-2.27 (m, 1H), 2.09-1.95 (m, 2H), 1.93-1.73 (m, 6H); LC/MS (APCI⁺) m/z 999 (M+H)⁺.

Example 38E: {[({(2S)-1,4-bis[2-(4-chloro-3-fluorophenoxy)acetamido]bicyclo[2.2.2]octan-2-yl}oxy)carbonyl]oxy}methyl[4-(phosphonooxy)phenyl]acetate

To a solution of Example 38D (0.102 g, 0.102 mmol) in ethyl acetate (4.0 mL) was added to 5% Pd/C (wet JM #9) (26.3 mg, 0.110 mmol) in a 20 mL RS10 Hast C reactor, which was then purged with argon. The mixture was stirred at 1200 rpm under 50 psi of hydrogen at 25° C., and vented after 6 minutes. The mixture was filtered through a filter funnel with a polyethylene frit packed with diatomaceous earth, concentrated under reduced pressure and dried under vacuum to give the title compound (70.0 mg, 0.086 mmol, 84%). ¹H NMR (501 MHz, DMSO-d₆) δ ppm 7.73 (s, 1H), 7.70 (s, 1H), 7.45 (td, J=8.9, 6.6 Hz, 2H), 7.20 (d, J=8.2 Hz, 2H), 7.10 (d, J=8.1 Hz, 2H), 6.99 (ddd, J=20.7, 11.4, 2.8 Hz, 2H), 6.79 (ddd, J=16.5, 8.9, 2.9 Hz, 2H), 5.71 (q, J=6.1 Hz, 2H), 5.36 (dd, J=9.4, 2.2 Hz, 1H), 4.49-4.38 (m, 4H), 3.71 (d, J=2.1 Hz, 2H), 2.49-2.41 (m, 1H), 2.38-2.28 (m, 1H), 2.06-1.71 (m, 8H); MS (ESI) m/z 815.1 (M−H)⁻.

The following compounds are prepared using methodologies described in the previous examples.

Example 39: (2S)-4-[2-(4-chloro-3-fluorophenoxy)acetamido]-1-[3-(4-chloro-3-fluorophenyl)propanamido]bicyclo[2.2.2]octan-2-yl 3-[2,4-dimethyl-3,6-bis(phosphonooxy)phenyl]-3-methylbutanoate (Compound 138) Example 40: (3-{3-[4-({(2S)-4-[2-(4-chloro-3-fluorophenoxy)acetamido]-1-[3-(4-chloro-3-fluorophenyl)propanamido]bicyclo[2.2.2]octan-2-yl}oxy)-2-methyl-4-oxobutan-2-yl]-2,6-dimethyl-4-(phosphonooxy)phenoxy}propyl)phosphonic Acid (Compound 139) Example 41: ({(2S)-4-[2-(4-chloro-3-fluorophenoxy)acetamido]-1-[3-(4-chloro-3-fluorophenyl)propanamido]bicyclo[2.2.2]octan-2-yl}oxy)methyl 3-[2,4-dimethyl-6-(phosphonooxy)phenyl]-3-methylbutanoate (Compound 140) Example 42: ({(2S)-4-[2-(4-chloro-3-fluorophenoxy)acetamido]-1-[3-(4-chloro-3-fluorophenyl)propanamido]bicyclo[2.2.2]octan-2-yl}oxy)methyl 3-[2,4-dimethyl-3,6-bis(phosphonooxy)phenyl]-3-methylbutanoate (Compound 141) Characterization Assays Abbreviations

D5W for 5% dextrose in water; DMSO for dimethyl sulfoxide; HPLC-MS/MS for high-performance liquid chromatography with tandem mass spectrometry; HPMC for hydroxypropyl methylcellulose; PEG for polyethylene glycol; rpm for revolutions per minute;

Example 43: Solubility Assay

Solubility measurements were performed using a high throughput 96-well plate based assay. For this, 0.2 mg of sample compound was weighed and added to a sample plate in triplicate and 650 μL of pH 7.4 phosphate buffer was added. The sample plate was capped, allowed to shake on a plate shaker for two days at room temperature, and then centrifuged (2000 rpm, 10 minutes) to settle undissolved solids. Approximately 600 μL of the aqueous supernatant slurry from each sample plate was transferred to a filter plate mounted on top of a receiver plate. Vacuum was applied to collect the filtrate. Appropriate dilutions were made in 50/50 methanol/water and quantification for concentration was performed against known standards by UV-UPLC. Solubilities of select compounds are reported in Table 2. Some preferred compounds of the invention have greater solubility than the parent compound (P), N,N′-[(2S)-2-hydroxybicyclo[2.2.2]octane-1,4-diyl]bis[2-(4-chloro-3-fluorophenoxy)acetamide].

TABLE 2 Solubility of exemplary compounds of the invention. Solubility Compound (pH 7.4 phosphate buffer) No. (μM) 100 <0.82 102 59 115 936 116 903 117 598 120 812 121 >1000 122 317 125 152 127 645 131 >1000 P¹ 0.00005 ¹P = N,N′-[(2S)-2-hydroxybicyclo[2.2.2]octane-1,4-diyl]bis[2-(4-chloro-3-fluorophenoxy)acetamide]

Example 44: Pharmacokinetic Assay

Six to eight week old CD₁ male mice were dosed with compounds orally at a dosing volume of 10 mL/kg in Vehicle A ((5:2:20:73 (v/v) dimethyl sulfoxide:Tween 80 (Polysorbate 80):PEG-400:D5W (5% dextrose in water)) or Vehicle B (0.5% hydroxypropyl methylcellulose with 1 equivalent of NaOH). Blood was drawn into ethylenediaminetetraacetic acid (EDTA) charged capillary tubes via the tail vein at the following timepoints: 0.25, 0.5, 1, 3, 6, 9, 12 and 24 h, N=3 measurements per timepoint (mice bled at each timepoint). Blood was centrifuged at 3000 rpm and plasma harvested. Terminal brain tissue was collected, flash frozen and analyzed for drug concentrations.

Plasma or tissue samples and standards were extracted by protein precipitation with 50:50 methanol: acetonitrile containing internal standards. The supernatant was diluted with water (for prodrugs that are potentially unstable in acid) or 0.1% formic acid in water before injection into an HPLC-MS/MS system for separation and quantitation. The analytes were separated from matrix components using reverse phase chromatography on a 30×2.1 mm 5 μm Fortis PACE C18 column using the following gradient (Mobile Phase A=25 mM ammonium bicarbonate and 25 mM ammonium hydroxide in water; Mobile Phase B=acetonitrile) at a flow rate of 1.5 mL/minute.

Total Flow A B Step Time(minute) Rate(μL/minute) (%) (%) 0 0.00 1500 95.0 5.0 1 0.20 1500 95.0 5.0 2 0.80 1500 5.0 95.0 3 1.00 1500 5.0 95.0 4 1.02 1500 95.0 5.0

The tandem mass spectrometry analysis was carried out on SCIEX™ triple quadrupole mass spectrometer with an electrospray ionization interface, in positive (or negative) ion mode. Data acquisition and evaluation were performed using Analyst® software (SCIEX™) Pharmacokinetic parameter values of select compounds are reported in Table 3. All pharmacokinetic parameters reported are for the parent compound, N,N-[(2S)-2-hydroxybicyclo[2.2.2]octane-1,4-diyl]bis[2-(4-chloro-3-fluorophenoxy)acetamide], following conversion after dosing the prodrug. Doses are reported for the parent drug equivalent (P). Preferred compounds of the invention have Cmax and AUC values greater than the parent compound.

TABLE 3 Pharmacokinetic parameter values of exemplary compounds of the invention following conversion to the parent compound after dosing the prodrug. Dose Compound (mg/kg) Cmax AUC PK No. equivalent (ng/mL) (ng*h/mL) vehicle 100 3 82 422  A¹ 101 3 394 1500 A 30 2620 13500 A 102 3 88 672 A 30 171 1310 A 103 3 260 1020 A 30 3200 14700 A 104 3 216 928 A 30 2270 9810 A 105 3 163 642 A 30 2710 9860 A 106 3 15 114 A 109 3 47 358 A 30 461 2430 A 110 3 163 499  B² 30 1410 6440 B 111 30 31 105 B 113 30 32 335 B 114 3 128 851 B 30 504 2550 B 115 3 19 145 B 30 79 962 B 116 3 4.3 10 B 30 723 2700 B 117 30 146 502 B 118 3 4.1 9.5 B 30 163 361 B 119 3 45 219 B 30 445 2650 B 120 3 90 544 B 30 148 916 B 121 3 37 242 B 30 1210 3849 B 122 30 14 26 B 125 30 5.3 36 B 126 30 0 0 B 127 3 35 166 B 30 1100 4530 B 131 3 101 515 B 30 661 5540 B 137 3 112 379 B 30 908 6050 B P⁴ 3 28 188  C³ 30 113 656 C ¹Vehicle A = 5:2 20:73 (v/v) DMSO:Tween 80 (Polysorbate 80):PEG-400:D5W (5% dextrose in water). ²Vehicle B = 0.5% HPMC with 1 equivalent NaOH. ³Vehicle C = 0.5% HPMC. ⁴P = N,N′-[(2S)-2-hydroxybicyclo[2.2.2]octane-1,4-diyl]bis[2-(4-chloro-3-fluorophenoxy)acetamide]

EQUIVALENTS AND SCOPE

In the claims articles such as “a,” “an,” and “the” may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The invention includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The invention includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process.

Furthermore, the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims are introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the invention, or aspects of the invention, is/are referred to as comprising particular elements and/or features, certain embodiments of the invention or aspects of the invention consist, or consist essentially of, such elements and/or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or subrange within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present invention that falls within the prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the invention can be excluded from any claim, for any reason, whether or not related to the existence of prior art.

Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention, as defined in the following claims. 

1. A compound represented by Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is selected from the group consisting of —C(O)—C₁₋₄alkyl, —C(O)—O—C₁₋₄alkyl, —C(O)—N(R^(a))—C₁₋₄alkyl, —C(O)—C₁₋₄alkylene-C₁₋₄alkoxy, —C(O)—C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, -methylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—O—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—N(R^(a))—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-P(O)(OH)₂, —C(O)—C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)(O—C₁₋₅alkylene-P(O)(OH)₂), -methylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂ or -methylene-O—C(O)C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—O—C₁₋₅alkylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, —C(O)—N(R^(a))-heteroarylene-C₁₋₂alkylene-O—P(O)(OH)₂, —P(O)(OH)₂, —SO₃H, —SO₂NR^(a)R^(b), —C(O)-heteroaryl, —C(O)—C₁₋₅alkylene-O—C₁₋₅alkylene-phenyl and methylene-C₁₋₅alkoxide; wherein —C(O)—C₁₋₄alkyl is substituted by one or two substituents each independently selected from the group consisting of NR^(a)R^(b) and —CO₂H; and wherein —C(O)—O—C₁₋₄alkyl, —C(O)—N(R^(a))—C₁₋₄alkyl, —C(O)—C₁₋₄alkylene-C₁₋₄alkoxy, —C(O)—C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy, methylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—O—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—N(R^(a))—C₁₋₅alkylene-O—P(O)(OH)₂, —C(O)—C₁₋₅alkylene-P(O)(OH)₂, —C(O)—C₁₋₅alkylene-phenylene-0-P(O)(OH)₂, —C(O)—C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)(O—C₁₋₅alkylene-P(O)(OH)₂), -methylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂ or -methylene-O—C(O)C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂, —C(O)—O—C₁₋₅alkylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, —C(O)-heteroaryl, —C(O)—C₁₋₅alkylene-O—C₁₋₅alkylene-phenyl and —C(O)—N(R^(a))-heteroarylene-C₁₋₂alkylene-O—P(O)(OH)₂ may optionally be substituted by one, two, three or four substituents each independently selected from the group consisting of halogen, —CO₂H, —NR^(a)R^(b) and C₁₋₂alkyl (optionally substituted by one, two or three fluorines) and aryl; and R^(a) and R^(b) are independently selected, for each occurrence, from the group consisting of hydrogen and C₁₋₃alkyl with the proviso that the compound is not

 or a pharmaceutically acceptable salt thereof.
 2. The compound of claim 1, wherein R¹ is —C(O)—C₁₋₄alkyl, wherein —C(O)—C₁₋₄alkyl is substituted by one or two substituents each independently selected from the group consisting of NR^(a)R^(b) and —CO₂H.
 3. The compound of claim 1, wherein R¹ is selected from the group consisting of


4. The compound of claim 1, wherein R¹ is —C(O)—O—C₁₋₄alkyl.
 5. The compound of claim 4, wherein R¹ is selected from the group consisting of


6. The compound of claim 1, wherein R¹ is —C(O)—N(R^(a))—C₁₋₄alkyl, wherein —C(O)—N(R^(a))—C₁₋₄alkyl may optionally be substituted by one or two CO₂H groups.
 7. The compound of claim 6, wherein R¹ is represented by


8. The compound of claim 1, wherein R¹ is —C(O)—C₁₋₄alkylene-C₁₋₄alkoxy.
 9. The compound of claim 8, wherein R¹ is selected from the group consisting of


10. The compound of claim 1, wherein R¹ is —C(O)—C₁₋₄alkylene-O—C₁₋₄alkylene-C₁₋₄alkoxy.
 11. The compound of claim 10, wherein R¹ is represented by

12.-13. (canceled)
 14. The compound of claim 1, wherein R¹ is —C(O)—C₁₋₅alkylene-O—P(O)(OH)₂.
 15. The compound of claim 14, wherein R¹ is selected from the group consisting of


16. The compound of claim 1, wherein R¹ is —C(O)—O—C₁₋₅alkylene-O—P(O)(OH)₂.
 17. The compound of claim 16, wherein R¹ is selected from the group consisting of


18. The compound of claim 1, wherein R¹ is —C(O)—N(R^(a))—C₁₋₅alkylene-O—P(O)(OH)₂.
 19. The compound of claim 18, wherein R¹ is selected from the group consisting of


20. The compound of claim 1, wherein R¹ is —C(O)—C₁₋₅alkylene-P(O)(OH)₂.
 21. The compound of claim 20, wherein R¹ is represented by


22. The compound of claim 1, wherein R¹ is —C(O)—C₁₋₅alkylene-phenylene-O—P(O)(OH)₂—C(O)—C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂ or —C(O)—C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)(O—C₁₋₅alkylene-P(O)(OH)₂).
 23. The compound of claim 22, wherein R¹ is selected from the group consisting of


24. The compound of claim 1, wherein R¹ is —C(O)—N(R^(a))-heteroarylene-C₁₋₂alkylene-O—P(O)(OH)₂.
 25. The compound of claim 24, wherein R¹ is represented by


26. The compound of claim 1, wherein R¹ is —C(O)-heteroaryl.
 27. The compound of claim 26, wherein R¹ is selected from the group consisting of


28. The compound of claim 1, wherein R¹ is —C(O)—C₁₋₅alkylene-O—C₁₋₅alkylene-phenyl.
 29. The compound of claim 28, wherein R¹ is represented by


30. The compound of claim 1, wherein R¹ is methylene-C₁₋₅alkoxide.
 31. The compound of claim 30, wherein R¹ is represented by


32. The compound of claim 1, wherein R¹ is —C(O)—O—C₁₋₅alkylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂, -methylene-O—C(O)C₁₋₅alkylene-phenylene-O—P(O)(OH)₂ or -methylene-O—C(O)C₁₋₅alkylene-phenylene-(O—P(O)(OH)₂)₂.
 33. The compound of claim 32, wherein R¹ is represented by


34. The compound of claim 1, wherein R¹ is selected from the group consisting of —P(O)(OH)₂, —SO₃H, and —SO₂NH₂.
 35. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
 36. A pharmaceutically acceptable composition comprising a compound of claim 35 and a pharmaceutically acceptable carrier.
 37. (canceled)
 38. A method of treating a neurodegenerative disease, a leukodystrophy, a cancer, an inflammatory disease, an autoimmune disease, a viral infection, a skin disease, a fibrotic disease, a hemoglobin disease, a kidney disease, a hearing loss condition, an ocular disease, a musculoskeletal disease, a metabolic disease, or a mitochondrial disease, or a disease related to a modulation of eIF2B activity or levels, eIF2α activity or levels, or the activity or levels of a component of the eIF2 pathway or the ISR pathway, in a patient in need thereof, comprising administering to the patient an effective amount of a compound of claim
 35. 39.-61. (canceled) 